LOCUS       sequence-271517-        4869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A cytoplasmic expression of a monomeric phytochrome-based 
            near-infrared fluorescent protein emiRFP703 (enhanced miRFP703).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4869)
  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 4869)
  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: pemiRFP703-N1
FEATURES             Location/Qualifiers
     source          1..4869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        346..549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     500..520
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      578..587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1638..1759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1675..1694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1729..1748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1766..2221)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1903..1924)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2115..2134
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2248..2352
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2354..2711
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2354..2545
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2377..2397)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2562..2697
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2746..3540
                     /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             2746..3540
                     /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(2800..2819)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3410..3429
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3728..3747)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3772..3819
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4148..4736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4637..4656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
       61 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      121 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      181 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      241 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      301 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      361 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      421 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      481 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      541 agcagagctg gtttagtgaa ccgtcagatc cgctagcgcc accatggcgg aaggctccgt
      601 cgccaggcag cctgacctct tgacctgcga acatgaagag atccacctcg ccggctcgat
      661 ccagccgcat ggcgcgcttc tggtcgtcag cgaacatgat catcgcgtca tccaggccag
      721 cgccaacgcc gcggaatttc tgaatctcgg aagcgtactc ggcgttccgc tcgccgagat
      781 cgacggcgat ctgttgatca agatcctgcc gcatctcgat cccaccgccg aaggcatgcc
      841 ggtcgcggtg cgctgccgga tcggcaatcc ctctacggag tactgcggtc tgatgcatcg
      901 gcctccggaa ggcgggctga tcatcgaact cgaacgtgcc ggcccgtcga tcgatctgtc
      961 aggcacgctg gcgccggcgc tggagcggat ccgcacggcg ggttcactgc gcgcgctgtg
     1021 cgatgacacc gtgctgctgt ttcagcagtg caccggctac gaccgggtga tggtgtatcg
     1081 tttcgatgag caaggccacg gcctggtatt ctccgagtgc catgtgcctg ggctcgaatc
     1141 ctatttcggc aaccgctatc cgtcgtcgct ggtcccgcag atggcgcggc agctgtacgt
     1201 gcggcagcgc gtccgcgtgc tggtcgacgt cacctatcag ccggtgccgc tggagccgcg
     1261 gctgtcgccg ctgaccgggc gcgatctcga catgtcgggc tgcttcctgc gctcgatgtc
     1321 gccgatccat ctgcagttcc tgaaggacat gggcgtgcgc gccaccctgg cggtgtcgct
     1381 ggtggtcggc ggcaagctgt ggggcctggt tgtctgtcac cattatctgc cgcgcttcat
     1441 ccgtttcgag ctgcgggcga tctgcaaacg gctcgccgaa aggatcgcga cgcggatcac
     1501 cgcgcttgag agctaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     1561 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1621 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1681 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1741 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1801 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1861 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1921 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1981 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2041 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2101 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2161 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2221 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2281 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2341 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2401 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2461 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2521 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2581 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2641 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2701 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2761 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2821 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2881 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     2941 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3001 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3061 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3121 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3181 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3241 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3301 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3361 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3421 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3481 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3541 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3601 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3661 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3721 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3781 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3841 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3901 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3961 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4021 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4261 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4801 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     4861 tagtaatca
//
