LOCUS       sequence-231170-        4960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric near-infrared fluorescent protein miRFP703 in mammalian 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4960)
  AUTHORS   Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, 
            Verkhusha VV
  TITLE     Bright monomeric near-infrared fluorescent proteins as tags and 
            biosensors for multiscale imaging.
  JOURNAL   Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405.
  PUBMED    27539380
REFERENCE   2  (bases 1 to 4960)
  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: "2016-08-19"; volume: "7:12405. doi"; pages: " 
            10.1038/ncomms12405"
COMMENT     Sequence Label: pmiRFP703-N1
FEATURES             Location/Qualifiers
     source          1..4960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        416..619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     570..590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    642..722
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      724..733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             730..1677
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    1799..1920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1836..1855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1890..1909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1927..2382)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2064..2085)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2276..2295
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2409..2513
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2515..2872
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2515..2706
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2538..2558)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2723..2858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2907..3701
                     /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             2907..3701
                     /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(2961..2980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3571..3590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3889..3908)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3933..3980
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4309..4897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4798..4817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
       61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      241 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      601 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
      661 gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
      721 gtcgccacca tggtagcagg tcatgcctct ggcagccccg cattcgggac cgcctctcat
      781 tcgaattgcg aacatgaaga gatccacctc gccggctcga tccagccgca tggcgcgctt
      841 ctggtcgtca gcgaacatga tcatcgcgtc atccaggcca gcgccaacgc cgcggaattt
      901 ctgaatctcg gaagcgtact cggcgttccg ctcgccgaga tcgacggcga tctgttgatc
      961 aagatcctgc cgcatctcga tcccaccgcc gaaggcatgc cggtcgcggt gcgctgccgg
     1021 atcggcaatc cctctacgga gtactgcggt ctgatgcatc ggcctccgga aggcgggctg
     1081 atcatcgaac tcgaacgtgc cggcccgtcg atcgatctgt caggcacgct ggcgccggcg
     1141 ctggagcgga tccgcacggc gggttcactg cgcgcgctgt gcgatgacac cgtgctgctg
     1201 tttcagcagt gcaccggcta cgaccgggtg atggtgtatc gtttcgatga gcaaggccac
     1261 ggcctggtat tctccgagtg ccatgtgcct gggctcgaat cctatttcgg caaccgctat
     1321 ccgtcgtcgc tggtcccgca gatggcgcgg cagctgtacg tgcggcagcg cgtccgcgtg
     1381 ctggtcgacg tcacctatca gccggtgccg ctggagccgc ggctgtcgcc gctgaccggg
     1441 cgcgatctcg acatgtcggg ctgcttcctg cgctcgatgt cgccgatcca tctgcagttc
     1501 ctgaaggaca tgggcgtgcg cgccaccctg gcggtgtcgc tggtggtcgg cggcaagctg
     1561 tggggcctgg ttgtctgtca ccattatctg ccgcgcttca tccgtttcga gctgcgggcg
     1621 atctgcaaac ggctcgccga aaggatcgcg acgcggatca ccgcgcttga gagctaagcg
     1681 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1741 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1801 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1861 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1921 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1981 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2041 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2101 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2161 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2221 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2281 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2341 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2401 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2461 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2521 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2581 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2641 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2701 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2761 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2821 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     2881 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2941 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3001 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3061 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3121 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3181 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3241 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3301 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3361 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3421 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3481 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3541 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3601 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3661 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3721 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3781 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3841 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     3901 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3961 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4021 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4081 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4141 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4201 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4261 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4321 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4381 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4441 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4501 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4561 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4621 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4681 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4741 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4801 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4861 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4921 acggttcctg gccttttgct ggccttttgc tcacatgttc
//
