LOCUS       sequence-250412-        5719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Keratin labeling with near-infrared fluorescent protein 
            miRFP670nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5719)
  AUTHORS   Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
  TITLE     Smallest near-infrared fluorescent protein evolved from 
            cyanobacteriochrome as versatile tag for spectral multiplexing.
  JOURNAL   Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
  PUBMED    30655515
REFERENCE   2  (bases 1 to 5719)
  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: "2019-01-17"; volume: "10(1):279. doi"; pages: " 
            10.1038/s41467-018-08050-8"
COMMENT     Sequence Label: pKeratin-miRFP670nano
FEATURES             Location/Qualifiers
     source          1..5719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2532..2653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2569..2588)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2623..2642
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2660..3115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2797..2818)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3009..3028
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3142..3246
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3248..3605
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3248..3439
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3271..3291)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3456..3591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3640..4434
                     /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             3640..4434
                     /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(3694..3713)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4304..4323
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4622..4641)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4666..4713
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5042..5630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5531..5550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact tttcaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag caccatgagc ttcaccactc gctccacctt ctccaccaac
      661 taccggtccc tgggctctgt ccaggcgccc agctacggcg cccggccggt cagcagcgcg
      721 gccagcgtct atgcaggcgc tgggggctct ggttcccgga tctccgtgtc ccgctccacc
      781 agcttcaggg gcggcatggg gtccgggggc ctggccaccg ggatagccgg gggtctggca
      841 ggaatgggag gcatccagaa cgagaaggag accatgcaaa gcctgaacga ccgcctggcc
      901 tcttacctgg acagagtgag gagcctggag accgagaacc ggaggctgga gagcaaaatc
      961 cgggagcact tggagaagaa gggaccccag gtcagagact ggagccatta cttcaagatc
     1021 atcgaggacc tgagggctca gatcttcgca aatactgtgg acaatgcccg catcgttctg
     1081 cagattgaca atgcccgtct tgctgctgat gactttagag tcaagtatga gacagagctg
     1141 gccatgcgcc agtctgtgga gaacgacatc catgggctcc gcaaggtcat tgatgacacc
     1201 aatatcacac gactgcagct ggagacagag atcgaggctc tcaaggagga gctgctcttc
     1261 atgaagaaga accacgaaga ggaagtaaaa ggcctacaag cccagattgc cagctctggg
     1321 ttgaccgtgg aggtagatgc ccccaaatct caggacctcg ccaagatcat ggcagacatc
     1381 cgggcccaat atgacgagct ggctcggaag aaccgagagg agctagacaa gtactggtct
     1441 cagcagattg aggagagcac cacagtgctc accacacagt ctgctgaggt tggagctgct
     1501 gagacgacgc tcacagagct gagacgtaca gtccagtcct tggagatcga cctggactcc
     1561 atgagaaatc tgaaggccag cttggagaac agcctgaggg aggtggaggc ccgctacgcc
     1621 ctacagatgg agcagctcaa cgggatcctg ctgcaccttg agtcagagct ggcacagacc
     1681 cgggcagagg gacagcgcca ggcccaggag tatgaggccc tgctgaacat caaggtcaag
     1741 ctggaggctg agatcgccac ctaccgccgc ctgctggaag atggcgagga ctttaatctt
     1801 ggtgatgcct tggacagcag caactccatg caaaccatcc aaaagaccac cacccgccgg
     1861 atagtggatg gcaaagtggt gtctgagacc aatgacacca aagttctgag gcatcgaatt
     1921 ctgcagtcga cggtacccat ggcaaacctg gacaagatgc tgaataccac agtaacagag
     1981 gtgcggcagt tcctgcaggt ggacagagtg tgcgtgttcc agtttgagga ggattatagc
     2041 ggagtggtgg tggtggaggc cgtggacgat aggtggatct ccatcctgaa gacccaggtg
     2101 cgggatagat acttcatgga gacaaggggc gaggagtatt ctcacggccg ctaccaggcc
     2161 atcgccgaca tctacaccgc aaacctgaca gagtgctaca gggatctgct gacacagttt
     2221 caggtgagag caatcctggc cgtgcccatc ctgcagggca agaagctgtg gggcctgttg
     2281 gtggcacacc agctggcggc ccctagacag tggcagacct gggagatcga ctttctgaag
     2341 cagcaggccg tggtggtggg catcgccatc cagcagagct agctgaattc tgcagatatc
     2401 cagcacagtg gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     2461 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2521 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2581 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2641 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2701 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2761 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2821 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2881 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2941 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3001 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3061 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     3121 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3181 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3241 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3301 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3421 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3481 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3541 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3601 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3661 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3721 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3781 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     3841 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3901 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3961 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4021 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4081 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4141 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     4201 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4261 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4321 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4381 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4441 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4501 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4561 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     4621 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4681 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4741 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4801 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4861 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     4921 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4981 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5041 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5101 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5161 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5221 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5281 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5341 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5401 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5461 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5521 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5581 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5641 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5701 cgttatcccc tgattctgt
//
