LOCUS       sequence-243906-        5717 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 5717)
  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 5717)
  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..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..250
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     201..221
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2187..2308
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2224..2243)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2278..2297
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2315..2770)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2452..2473)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2664..2683
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2797..2901
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2903..3260
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2903..3094
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2926..2946)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3111..3246
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3295..4089
                     /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             3295..4089
                     /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(3349..3368)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3959..3978
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4277..4296)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4321..4368
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4697..5285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5186..5205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5460..5717,1..46)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
       61 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      121 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      181 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      241 aagcagagct ggtttagtga accgtcagat ccgctagcac catgagcttc accactcgct
      301 ccaccttctc caccaactac cggtccctgg gctctgtcca ggcgcccagc tacggcgccc
      361 ggccggtcag cagcgcggcc agcgtctatg caggcgctgg gggctctggt tcccggatct
      421 ccgtgtcccg ctccaccagc ttcaggggcg gcatggggtc cgggggcctg gccaccggga
      481 tagccggggg tctggcagga atgggaggca tccagaacga gaaggagacc atgcaaagcc
      541 tgaacgaccg cctggcctct tacctggaca gagtgaggag cctggagacc gagaaccgga
      601 ggctggagag caaaatccgg gagcacttgg agaagaaggg accccaggtc agagactgga
      661 gccattactt caagatcatc gaggacctga gggctcagat cttcgcaaat actgtggaca
      721 atgcccgcat cgttctgcag attgacaatg cccgtcttgc tgctgatgac tttagagtca
      781 agtatgagac agagctggcc atgcgccagt ctgtggagaa cgacatccat gggctccgca
      841 aggtcattga tgacaccaat atcacacgac tgcagctgga gacagagatc gaggctctca
      901 aggaggagct gctcttcatg aagaagaacc acgaagagga agtaaaaggc ctacaagccc
      961 agattgccag ctctgggttg accgtggagg tagatgcccc caaatctcag gacctcgcca
     1021 agatcatggc agacatccgg gcccaatatg acgagctggc tcggaagaac cgagaggagc
     1081 tagacaagta ctggtctcag cagattgagg agagcaccac agtggtcacc acacagtctg
     1141 ctgaggttgg agctgctgag acgacgctca cagagctgag acgtacagtc cagtccttgg
     1201 agatcgacct ggactccatg agaaatctga aggccagctt ggagaacagc ctgagggagg
     1261 tggaggcccg ctacgcccta cagatggagc agctcaacgg gatcctgctg caccttgagt
     1321 cagagctggc acagacccgg gcagagggac agcgccaggc ccaggagtat gaggccctgc
     1381 tgaacatcaa ggtcaagctg gaggctgaga tcgccaccta ccgccgcctg ctggaagatg
     1441 gcgaggactt taatcttggt gatgccttgg acagcagcaa ctccatgcaa accatccaaa
     1501 agaccaccac ccgccggata gtggatggca aagtggtgtc tgagaccaat gacaccaaag
     1561 ttctgaggca tcgaattctg cagtcgacgg tacccatggc aaacctggac aagatgctga
     1621 ataccacagt aacagaggtg cggcagttcc tgcaggtgga cagagtgtgc gtgttccagt
     1681 ttgaggagga ttatagcgga gtggtggtgg tggaggccgt ggacgatagg tggatctcca
     1741 tcctgaagac ccaggtgcgg gatagatact tcatggagac aaggggcgag gagtattctc
     1801 acggccgcta ccaggccatc gccgacatct acaccgcaaa cctgacagag tgctacaggg
     1861 atctgctgac acagtttcag gtgagagcaa tcctggccgt gcccatcctg cagggcaaga
     1921 agctgtgggg cctgttggtg gcacaccagc tggcggcccc tagacagtgg cagacctggg
     1981 agatcgactt tctgaagcag caggccgtgg tggtgggcat cgccatccag cagagctagg
     2041 aattctgcag atatccagca cagtggcggc cgcgactcta gatcataatc agccatacca
     2101 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2161 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2221 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2281 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     2341 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2401 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2461 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2521 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2581 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2641 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2701 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2761 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2821 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2881 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2941 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3001 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3121 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3181 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3241 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3301 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3361 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3421 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3481 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3541 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3601 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3661 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3721 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3781 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3841 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3901 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3961 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4021 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4081 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4141 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4201 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4261 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4321 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4381 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4441 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4501 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4561 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4621 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4681 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4741 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4801 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4861 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4921 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4981 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5041 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5101 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5161 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5221 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5281 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5341 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     5401 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5461 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5521 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5581 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5641 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5701 atgaccttat gggactt
//
