Fluorescent Protein Guide: Empty Backbones
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Fluorescent proteins are genetically encoded tools that are used extensively by life scientists. The original green fluorescent protein (GFP) was cloned in 1992 (Prasher et al., Gene, 1992), and since then scientists have engineered numerous GFP-variants and non-GFP proteins that result in a diverse set of colors.
Addgene has assembled a collection of empty plasmid backbones with different fluorescent tags for you to create fusion proteins with your gene of interest. These are organized By Color of the fluorescent protein and By Expression Species .
The content for this page was generated with the help of Erik Snapp .
Empty Backbones for Fluorescent Protein Fusions (Organized by Color)
For each table:
- Excitation and Emission maximum are listed in nm.
- Brightness is the product of exctinction coefficient and quantum yield divided by 1000.
- pKa is the pH at which fluorescence intensity is 50% of its maximum value.
- Maturation is the time for fluorescence to obtain half-maximal value.
- Structure describes the monomeric or oligomeric state of the protein. Many FPs are monomers at low concentration but are prone to dimerization at high concentration, so this is noted here. Proteins that are derived from GFP and contain the A206K mutation are monomeric at all concentrations, so this is mutation is noted when present.
- For more information on the properties of many of these proteins, see:
- Cranfill et al., Nature Methods 2016 Jul;13(7):557-62.
- FPbase: A database for users of fluorescent proteins created by Talley Lambert at Harvard Medical School
- More References below
Jump to:
Blue/UV
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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Sirius | 355 | 424 | 4 | 3.0 | Prone to dimerization |
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SBFP2 | 380 | 446 | 13 | Monomer (A206K) |
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Azurite | 383 | 450 | 14 | 5 | Prone to dimerization |
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mAzurite | 383 | 450 | 14 | 5 | Monomer (A206K) |
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EBFP2 | 383 | 448 | 17 | 4.5 | Prone to dimerization |
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moxBFP | 385 | 448 | 17 | Monomer (A206K) |
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mKalama1 | 385 | 456 | 16 | 5.5 | Monomer (A206K) |
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mTagBFP2 | 399 | 456 | 32 | 2.7 | 12 min | Prone to dimerization |
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Cyan
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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Aquamarine | 420 | 474 | 23 | 3.3 | Prone to dimerization |
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ECFP | 433 | 475 | 13 | 4.7 | Prone to dimerization |
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Cerulean | 433 | 475 | 27 | 4.7 | Monomer (A206K) |
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mCerulean3 | 433 | 475 | 32 | 3.2 | Monomer (A206K) |
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moxCerulean3 | 434 | 474 | 36 | 3.2 | Monomer (A206K) |
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SCFP3A | 433 | 474 | 17 | 4.5 | Monomer (A206K) |
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mTurquoise2 | 434 | 474 | 28 | 3.1 | Monomer (A206K) |
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CyPet | 435 | 477 | 18 | 5 | Monomer |
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AmCyan1 | 453 | 486 | 11 | Tetramer |
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MiCy (Midoriishi-Cyan) | 472 | 495 | 25 | 6.6 | Dimer |
Green
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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iLOV | 447 | 497 | Monomer | ||||
AcGFP1 | 475 | 505 | 28 | Prone to dimerization | |||
sfGFP | 485 | 507 | 54 | Prone to dimerization |
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moxGFP | 486 | 510 | 50 | Monomer (A206K) |
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mEmerald | 487 | 509 | 39 | 6 | Monomer (A206K) |
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EGFP | 488 | 507 | 34 | 6 | Prone to dimerization |
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mEGFP | 488 | 507 | 34 | 6 | Monomer (A206K) |
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mAzamiGreen | 492 | 505 | 41 | 5.8 | Monomer |
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cfSGFP2 | 493 | 517 | 34 | 5.4 | Monomer (A206K) |
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ZsGreen | 493 | 505 | 39 | Tetramer |
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SGFP2 | 495 | 512 | 30 | Monomer (A206K) |
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Clover | 505 | 515 | 84 | 6.1 | 30 min | Prone to dimerization |
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dClover2 | 505 | 515 | Dimer (A206K) |
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mClover3 | 506 | 518 | 85 | 6.5 | Monomer (A206K) |
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mNeonGreen | 506 | 517 | 93 | 5.7 | 10 min | Monomer |
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Yellow
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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EYFP | 513 | 527 | 51 | 6.9 | Prone to dimerization |
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Topaz | 514 | 527 | 57 | Prone to dimerization |
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mTopaz | 514 | 527 | Monomer (A206K) |
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mVenus | 515 | 528 | 53 | 6 | Monomer (A206K) |
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moxVenus | 514 | 526 | 44 | Monomer (A206K) |
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SYFP2 | 515 | 527 | 69 | 6 | Monomer (A206K) |
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mGold | 515 | 530 | 68 | 5.9 | Monomer |
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mCitrine | 516 | 529 | 59 | 5.7 | Monomer (A206K) |
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YPet | 517 | 530 | 80 | 5.6 | Prone to dimerization |
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ZsYellow1 | 529 | 539 | 8 | Tetramer |
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mPapaya1 | 530 | 541 | 36 | 6.8 | Monomer |
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mCyRFP1 | 528 | 594 | 18 | 5.6 | Monomer |
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Orange
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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mKusabira-Orange (mKO) | 548 | 559 | 31 | 5 | 4.5 hr | Monomer |
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mOrange | 548 | 562 | 49 | 6.5 | 2.5 hr | Monomer |
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mOrange2 | 549 | 565 | 35 | 6.5 | 4.5 hr | Monomer |
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mKO2 | 551 | 565 | 40 | 5.5 | Monomer | ||
TurboRFP | 553 | 574 | 62 | 4.4 | Dimer |
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tdTomato | 554 | 581 | 95 | 4.7 | 1 hr | Tandem-dimer |
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Red
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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mScarlet-H | 551 | 592 | 15 | 4.8 | 4.4 hr | Monomer |
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mNectarine | 558 | 578 | 26 | 6.9 | 30 min | Monomer |
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mRuby2 | 559 | 600 | 43 | 5.3 | 150 min | Prone to dimerization |
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eqFP611 | 559 | 611 | 35 | Tetramer |
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DsRed2 | 563 | 582 | 24 | Tetramer |
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mApple | 568 | 592 | 37 | 6.5 | 30 min | Monomer |
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mScarlet | 569 | 594 | 71 | 5.3 | 2.9 hr | Monomer |
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mScarlet-I | 569 | 593 | 57 | 5.4 | 0.6 hr | Monomer |
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mStrawberry | 574 | 596 | 26 | 4.5 | 50 min | Monomer |
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FusionRed | 580 | 608 | 18 | Monomer |
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mRFP1 | 584 | 607 | 13 | 4.5 | 1 hr | Monomer |
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mCherry | 587 | 610 | 16 | 4.5 | 15 min | Monomer |
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mCherry2 | 587 | 610 | Monomer |
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Far-Red
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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mCrimson3 | 588 | 615 | - | Monomer |
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HcRed1 | 588 | 618 | 0.3 | Dimer |
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dKatushka | 588 | 635 | 22 | 5.5 | Dimer |
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mKate1.3 | 588 | 635 | 25 | Monomer |
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mPlum | 590 | 649 | 4 | 4.5 | 100 min | Monomer |
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mRaspberry | 598 | 625 | 13 | 55 min | Monomer |
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TagRFP675 | 598 | 675 | 4 | 5 | Prone to dimerization |
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mNeptune | 600 | 650 | 13 | 5.4 | 35 min | Prone to dimerization |
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mNeptune2 | 600 | 650 | 21 | Prone to dimerization |
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mCarmine | 603 | 675 | 5.81 | 5.58 | Monomer |
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mCardinal | 604 | 659 | 17 | 5.3 | 27 min | Prone to dimerization |
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mMaroon | 610 | 652 | - | Monomer |
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TagRFP657 | 611 | 657 | 0.34 | 5.3 | 27 min | Prone to dimerization |
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smURFP + BV | 642 | 670 | 32 | 39 min | Prone to dimerization |
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Near-Infrared
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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miRFP670 | 642 | 670 | 12 | 4.5 | Monomer |
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iRFP670 | 643 | 670 | 13 | 4 | ~5 hr | Dimer |
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iRFP682 | 663 | 682 | 10 | 4.5 | ~5hr | Dimer |
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miRFP703 | 673 | 703 | 8 | 4.5 | Monomer |
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iRFP702 | 673 | 702 | 8 | 4.5 | ~5hr | Dimer |
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miRFP709 | 683 | 709 | 4 | 4.5 | Monomer |
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mIFP | 683 | 704 | 7 | 3.5 | Monomer | ||
IFP2.0 | 684 | 708 | 7 | Prone to dimerization | |||
iRFP (aka iRFP713) | 690 | 713 | 6 | 4.5 | 2.8 hr | Dimer |
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iSplit | 690 | 713 | 5 | Dimer |
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iRFP720 | 702 | 720 | 6 | 4.5 | ~5hr | Dimer |
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Long Stokes Shift
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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T-sapphire | 399 | 511 | 26 | 4.9 | 1.3 hr | Prone to dimerization |
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mT-sapphire | 399 | 511 | Monomer (A206K) |
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mAmetrine | 406 | 526 | 26 | 6 | 0.8 hr | Monomer (A206K) |
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LSSmOrange | 437 | 572 | 23 | 5.7 | 2.3 hr | Monomer |
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mKeima Red | 440 | 620 | 3 | 6.5 | 4.4 hr | Monomer |
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dKeima Red | 440 | 616 | 22 | 6.5 | Dimer |
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LSSmKate1 | 463 | 624 | 3 | 3.2 | 1.7 hr | Monomer |
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LSSmKate2 | 460 | 605 | 4 | 2.7 | 2.5 hr | Monomer |
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Photoactivatable (e.g. off to on)
Excitation and Emission wavelengths after activation.
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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Phamret | 458 | 517 | 14 | Monomer |
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PA-sfGFP | 485 | 511 | Prone to dimerization |
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mPA-Emerald | 487 | 509 | - | Monomer (A206K) |
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PA-GFP | 504 | 517 | 14 | Monomer (A206K) |
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PATagRFP | 562 | 595 | 25 | 5.3 | 75 min | Prone to dimerization |
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PAmCherry1 | 564 | 595 | 8 | 6.3 | 23 min | Monomer |
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PAmCherry2 | 570 | 596 | 13 | 6.2 | 18 min | Monomer |
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PAmCherry3 | 570 | 596 | 5 | 6.2 | 25 min | Monomer |
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PAmKate | 568 | 628 | 5 | 5.6 | 19 min | Monomer |
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PAiRFP1 | 690 | 717 | 3 | Dimer |
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PAiRFP2 | 692 | 719 | 3 | Dimer |
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Photoconvertible (e.g. green to red)
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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Dendra2 | 490 / 553 | 507 / 573 | 23/19 | 6.6/6.9 | Monomer |
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mEos2 | 506 / 571 | 516 / 581 | 47 / 30 | 5.6/6.4 | Prone to oligomerization |
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mEos3.2 | 507 / 572 | 516 / 580 | 53 / 18 | 5.4/5.8 | 20 min | Monomer |
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mEos4a | 505 / 569 | 516 / 581 | Prone to oligomerization | ||||
mEos4b | 505 / 569 | 516 / 581 | Monomer | ||||
dEos | 505 / 569 | 516 / 581 | 55 / 20 | Dimer |
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tdEos | 505 / 569 | 516 / 581 | 55 / 20 | Tandem dimer | |||
mKikGR | 507 / 583 | 517 / 593 | 34 / 18 | 6.6/5.2 | Monomer | ||
Kaede | 508 / 572 | 518 / 580 | 87 / 20 | 5.6/5.6 | Tetramer | ||
PSmOrange | 548 / 634 | 565 / 662 | 58 / 9 | 6.2/5.6 | 1.6 hr | Monomer |
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PSmOrange2 | 546 / 619 | 561 / 651 | 31 / 7 | 6.6/5.4 | 3.5 hr | Monomer |
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Photoswitchable (e.g. off to on to off)
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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rsTagRFP | 440 / 567 | 585 / 585 | 0.02 / 4.0 | 6.6 | 43 min | Prone to dimerization |
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rsEmerald | 491 | 508 |
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rsGFP1 | 493 | 510 |
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Dronpa3 | 503 | 518 | 19 | Monomer |
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mGeos-M | 503 / 569 | 514 / 581 | 44 | 4.7 | Prone to dimerization |
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Fluorescent Timers
Protein | Excitation (nm) | Emission (nm) | Brightness | pKa | Maturation | Structure | Plasmids |
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Fast-FT | 403(blue), 583(red) | 466(blue), 606(red) | 14.9(blue), 6.8(red) | 2.8/4.1 | 7.1 hr (red) | Monomer |
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Medium-FT | 401(blue), 579(red) | 464(blue), 600(red) | 18.4(blue), 5.8(red) | 2.7/4.7 | 3.9 hr (red) | Monomer |
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Slow-FT | 402(blue), 583(red) | 465(blue), 604(red) | 11.7(blue), 4.2(red) | 2.6/4.6 | 28 hr (red) | Monomer |
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Empty Backbones for Fluorescent Protein Fusions (Organized by Expression Species)
Organism/Expression System | Plasmids |
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Mammalian |
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Insect |
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Zebrafish, Sea urchin, Xenopus, and C. elegans |
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Yeast |
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Bacteria |
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References:
- Cranfill et al.: Nature Methods July 2016, Vol. 13(7):557-62
- Olympus Confocal: The Fluorescent Protein Color Palette
- NIC Wiki: Nikon Imaging Center at UCSF / QB3 Wiki
- FPbase: A database created by Talley Lambert at Harvard Medical School
- Zeiss Microscopy: Optical Highlighters
- NIC Wiki PSFP: Nikon Imaging Center at UCSF / QB3 Wiki
- Stepanenko et al.: BioTechniques, Vol. 51, No. 5, November 2011, pp. 313–327
- Scholarpedia: Entry by Robert Campbell
- Spectra Database: Graphing site hosted at the University of Arizona
- Shaner et al.: Journal of Cell Science, Vol. 120, December 2007, pp. 4247-4260
- Shaner et al.: Nature Methods, Vol.5 No. 6, June 2008, pp. 545-51
- Shcherbakova et al.: Journal of the American Chemical Society, May 2012, Vol. 134 No.18, pp. 7913–7923
- Hoi et al.: Chemistry & Biology, October 2013, Vol. 20 No. 10, pp.1296-304
- Kogure et al.: Nature Biotechnology, May 2006, Vol. 24 No. 5, pp.577-81
- Shcherbo et al.: Nature Methods, September 2007, Vol. 4 No. 9, pp. 741-6
- Shemiakina et al.: Nature Communications, November 2012, Vol. 3 No. 1204
- Wiedenmann et al.: Journal of Biomedical Optics, Feburary 2005, Vol. 10 No. 1, p. 14003
- Chu et al.: Nature Methods, May 2014 Vol. 11 No. 5, pp.572-8
- Yu et al.: Nature Communications, May 2014 Vol. 15 No. 5, p. 3626
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