LOCUS       sequence-396432-        4937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a phytochrome-based near-infrared fluorescent protein, also known as
            iRFP713, used for in vivo imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4937)
  AUTHORS   Filonov GS, Piatkevich KD, Ting LM, Zhang J, Kim K, Verkhusha VV
  TITLE     Bright and stable near-infrared fluorescent protein for in vivo 
            imaging.
  JOURNAL   Nat Biotechnol. 2011 Jul 17. doi: 10.1038/nbt.1918.
  PUBMED    21765402
REFERENCE   2  (bases 1 to 4937)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2011 Jul 17. doi: 10.1038/nbt.1918."
COMMENT     Sequence Label: piRFP
FEATURES             Location/Qualifiers
     source          1..4937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..337
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     294..314
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(339..443)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      470..925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(557..576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     767..788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    932..1053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(943..962)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     997..1016
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1204..2154)
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     regulatory      2151..2160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2210..2413)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2239..2259)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2414..2717
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2892..3480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2972..2991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3809..3856
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3881..3900
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4088..4882)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4088..4882)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4199..4218)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4809..4828
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4917..4937,1..337))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4931..4937,1..129)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
       61 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
      121 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
      181 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
      241 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
      301 ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat
      361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
      421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
      481 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
      541 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
      601 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
      661 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
      721 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
      781 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
      841 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
      901 aacaaaatat taacgcttac aatttacgcg ttaagataca ttgatgagtt tggacaaacc
      961 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1021 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1081 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1141 ggtatggctg attatgatca gttatctaga tccggtggat cccgggcccg cggtaccgtc
     1201 gactcactct tccatcacgc cgatctgcca ggccagaacc tgggcgacta gctcgcaggc
     1261 ttggcggcca tcgagatcga cgtagtacgg cgttcggtga tggcaaacga tcaatcccca
     1321 cagtcgctcg ccgcgcaaaa tcgagatcga catcgtgccg tgcatgccta tgttgcgcat
     1381 gaattccaga tggacgggcg agacgctgcg caggatggcg aagctaagat caatcggccg
     1441 cccggtgacc ggattgaggt ctggggtgac cggcaccggc cgataattga tatcgggaat
     1501 gatccgtacc gggttgatgg tatagagccg acgggcctgc gccggcacgg ttgaggcagg
     1561 atagtgcagg cctagttttg actcgacctc ggcgcaccga tcctctgcga tcacttcgcc
     1621 gctgaagtcg gaggcgaagc gatagatcat cacccgatcg aagccggtaa tcttccgcac
     1681 ctcttgcgcc gcggcggcgc aggcgctttc caaggtttcg gcggcctgca ggcggcggat
     1741 ggcgctgttg gtgcggcgga agaacgcctg cggctcggcg acgtcccgct ggggaggctc
     1801 gagctcgagg aagatgagct gatcatggcg atgccaggag ccgatgaagc ctgcgtcctt
     1861 tcgcatcgtg aagccgacag tgatcggtgc tccgacggcc gccccgggct cggccaaggc
     1921 gatcgtcaga cggttgtgcg tctccgagtc gaagacatcg gccgcagagc ggccgatcag
     1981 ggcgccgatc gccagtccgg tgagttcggg aaggttgtcg ctgccggcaa cgatcgtcat
     2041 gtcggcggcg agggcgagca gcagtccatg cggttggatg gcaccgggga tatggatcgg
     2101 ctcatcgtcg caggtcaaga ggtcaggctg cctggcgacg gatccttcag ccatggtggc
     2161 ggcagatctg agtccggtag cgctagcgga tctgacggtt cactaaacca gctctgctta
     2221 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     2281 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     2341 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     2401 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     2461 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     2521 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     2581 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     2641 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     2701 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     2761 attactatta ataactaatg catggcggta atacggttat ccacagaatc aggggataac
     2821 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3061 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3121 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3181 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3241 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3301 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3361 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3421 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3481 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3541 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3601 tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc
     3661 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa
     3721 aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc
     3781 cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc
     3841 tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca
     3901 gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     3961 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     4021 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     4081 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     4141 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     4201 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     4261 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     4321 gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg
     4381 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     4441 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     4501 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     4561 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     4621 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     4681 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     4741 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     4801 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     4861 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg
     4921 caaaagccta ggcctcc
//
