LOCUS       sequence-150030-        4960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric near-infrared fluorescent protein miRFP709 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: pmiRFP709-N1
FEATURES             Location/Qualifiers
     source          1..4960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             13..960
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 709 nm (Shcherbakova et al., 2016)"
                     /label=miRFP709
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSFIPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    1082..1203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1119..1138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1173..1192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1210..1665)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1347..1368)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1559..1578
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1692..1796
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1798..2155
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1798..1989
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1821..1841)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2006..2141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2190..2984
                     /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             2190..2984
                     /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(2244..2263)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2854..2873
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3172..3191)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3216..3263
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3592..4180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4081..4100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4355..4658
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4659..4862
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4813..4833
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(4885..4960,1..5)
                     /label=MCS
                     /note="multiple cloning site"
ORIGIN
        1 ccggtcgcca ccatggtagc aggtcatgcc tctggcagcc ccgcattcgg gaccgcctct
       61 cattcgaatt gcgaacatga agagatccac ctcgccggct cgatccagcc gcatggcgcg
      121 cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg ccagcgccaa cgccgcggaa
      181 tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg agatcgacgg cgatctgttg
      241 atcaagatcc tgccgcatct cgatcccacc gccgaaggca tgccggtcgc ggtgcgctgc
      301 cggatcggca atccctctac ggagtactgc ggtctgatgc atcggcctcc ggaaggcggg
      361 ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc tgtcaggcac gctggcgccg
      421 gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc tgtgcgatga caccgtgctg
      481 ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt atcgtttcga tgagcaaggc
      541 cacggcctgg tattctccga gtgccatgtg cctgggctcg aatcctattt cggcaaccgc
      601 tatccgtcgt cgttcatccc gcagatggcg cggcagctgt acgtgcggca gcgcgtccgc
      661 gtgctggtcg acgtcaccta tcagccggtg ccgctggagc cgcggctgtc gccgctgacc
      721 gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga tgtcgccgat ccatctgcag
      781 ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt cgctggtggt cggcggcaag
      841 ctgtggggcc tggttgtctg tcaccattat ctgccgcgct tcatccgttt cgagctgcgg
      901 gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga tcaccgcgct tgagagctaa
      961 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1021 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1081 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1141 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1201 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1261 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1321 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1381 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     1441 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     1501 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     1561 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     1621 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     1681 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1741 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     1801 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     1861 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     1921 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1981 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2041 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2101 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     2161 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     2221 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     2281 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     2341 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     2401 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     2461 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     2521 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2581 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2641 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2701 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     2761 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     2821 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     2881 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     2941 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3001 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3061 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3121 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     3181 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3241 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3301 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3361 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3421 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     3481 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3541 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3601 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3661 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3721 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3781 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3841 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3901 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3961 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4021 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4081 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4141 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4201 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4261 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     4321 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4381 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4441 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4501 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4561 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4621 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     4681 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4741 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4801 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     4861 ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct
     4921 tcgaattctg cagtcgacgg taccgcgggc ccgggatcca
//
