LOCUS       sequence-271518-        4887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A cytoplasmic expression of a monomeric phytochrome-based 
            near-infrared fluorescent protein miRFP713.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4887)
  AUTHORS   Matlashov ME, Shcherbakova DM, Alvelid J, Baloban M, Pennacchietti 
            F, Shemetov AA, Testa I, Verkhusha VV
  TITLE     A set of monomeric near-infrared fluorescent proteins for multicolor
            imaging across scales.
  JOURNAL   Nat Commun. 2020 Jan 13;11(1):239. doi: 10.1038/s41467-019-13897-6.
  PUBMED    31932632
REFERENCE   2  (bases 1 to 4887)
  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 
            Commun."; date: "2020-01-13"; volume: "11(1):239. doi"; pages: " 
            10.1038/s41467-019-13897-6"
COMMENT     Sequence Label: pmiRFP713-N1
FEATURES             Location/Qualifiers
     source          1..4887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        28..331
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        332..535
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     486..506
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      564..573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             570..1520
                     /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
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     polyA_signal    1642..1763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1679..1698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1733..1752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1770..2225)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1907..1928)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2119..2138
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2252..2356
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2358..2715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2358..2549
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2381..2401)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2566..2701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2750..3544
                     /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             2750..3544
                     /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(2804..2823)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3414..3433
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3732..3751)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3776..3823
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4152..4740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4641..4660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
       61 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      121 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      181 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      241 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      301 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      361 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      421 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      481 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      541 agtgaaccgt cagatccgct agcgccacca tggcggaagg ctccgtcgcc aggcagcctg
      601 acctcttgac ctgcgacgat gagccgatcc atatccccgg tgccatccaa ccgcatggac
      661 tgctgctcgc cctcgccgcc gacatgacga tcgttgccgg cagcgacaac cttcccgaac
      721 tcaccggact ggcgatcggc gccctgatcg gccgctctgc ggccgatgtc ttcgactcgg
      781 agacgcacaa ccgtctgacg atcgccttgg ccgagcccgg ggcggccgtc ggagcaccga
      841 tcactgtcgg cttcacgatg cgaaaggacg caggcttcat cggctcctgg catcgccatg
      901 atcagctcat cttcctcgag ctcgagcctc cccagcggga cgtcgccgag ccgcaggcgt
      961 tcttccgccg caccaacagc gccatccgcc gcctgcaggc cgccgaaacc ttggaaagcg
     1021 cctgcgccgc cgcggcgcaa gaggtgcgga agattaccgg cttcgatcgg gtgatgatct
     1081 atcgcttcgc ctccgacttc agcggcgaag tgatcgcaga ggatcggtgc gccgaggtcg
     1141 agtcaaaact aggcctgcac tatcctgcct caaccgtgcc ggcgcaggcc cgtcggctct
     1201 ataccatcaa cccggtacgg atcattcccg atatcaatta tcggccggtg ccggtcaccc
     1261 cagacctcaa tccggtcacc gggcggccga ttgatcttag cttcgccatc ctgcgcagcg
     1321 tctcgcccgt ccatctggag ttcatgcgca acataggcat gcacggcacg atgtcgatct
     1381 cgattttgcg cggcgagcga ctgtggggat tgatcgtttg ccatcaccga acgccgtact
     1441 acgtcgatct cgatggccgc caagcctgca agagggtcgc cgagaggctg gccactcaga
     1501 tcggcgtgat ggaagagtga gcggccgcga ctctagatca taatcagcca taccacattt
     1561 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     1621 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1681 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1741 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     1801 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1861 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1921 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1981 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2041 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2101 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2161 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2221 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2281 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2341 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     2401 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2461 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2521 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2581 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2641 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2701 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2761 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2821 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2881 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     2941 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3001 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3061 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3121 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3181 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3241 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     3301 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3361 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3421 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3481 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3541 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3601 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3661 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     3721 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3781 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3841 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3901 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3961 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4021 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4081 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4141 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4201 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4261 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4321 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4381 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4441 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4501 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4561 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4621 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4681 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4741 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4801 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     4861 ttaatagtaa tcaattacgg ggtcatt
//
