LOCUS       sequence-204249-        5802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Photoconvertable cytoskeletal protein actin to study kinetics or 
            dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5802)
  AUTHORS   Doyle AD, Kutys ML, Conti MA, Matsumoto K, Adelstein RS, Yamada KM
  TITLE     Micro-environmental control of cell migration--myosin IIA is 
            required for efficient migration in fibrillar environments through 
            control of cell adhesion dynamics.
  JOURNAL   J Cell Sci. 2012 May 1;125(Pt 9):2244-56. doi: 10.1242/jcs.098806. 
            Epub 2012 Feb 10.
  PUBMED    22328520
REFERENCE   2  (bases 1 to 5802)
  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: "J Cell 
            Sci."; date: "2012-05-1"; volume: "125(Pt 9):2244-56. doi"; pages: "
            10.1242/jcs.098806. Epub 2012 Feb 10"
COMMENT     Sequence Label: pmKikGR Actin
FEATURES             Location/Qualifiers
     source          1..5802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     502..521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        776..1079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1080..1283
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1234..1254
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1328..2023
                     /codon_start=1
                     /product="monomeric photoconvertible Kikume green-to-red 
                     fluorescent protein"
                     /label=mKikGR1
                     /translation="MSVITSEMKIELRMEGSVNGHKFVITGKGSGRPYEGTQTVDLTVI
                     EGGPLPFAFDILTTAFHYGNRVFVEYPEEIVDYFKQSFPEGYSWERSMSYEDGGICLAT
                     NNITMKKDGSNTFVNEIRFDGTNFPANGPVMQRKTVKWEPSTEKMYVRDGVLKGDVEMA
                     LLLEGGGHYRCDFRTTYKAKKVVQLPDYHYVDHQMEITSHDKDYNKVKAYEHAKAYSGT
                     YRGAKYEFEA"
     polyA_signal    3305..3426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3342..3361)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3396..3415
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3433..3888)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3570..3591)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3782..3801
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3915..4019
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4021..4378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4021..4212
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4044..4064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4229..4364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4413..5207
                     /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             4413..5207
                     /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(4467..4486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5077..5096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5395..5414)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5439..5486
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
       61 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
      721 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      781 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      841 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      901 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      961 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1021 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1081 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1141 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1201 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1261 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt
     1321 cgccaccatg agtgtgatta catcagaaat gaagatcgag ctgcgtatgg aaggctctgt
     1381 aaacgggcac aagttcgtga ttacagggaa aggaagtggc agaccttacg agggaacaca
     1441 gactgtggac ctgacagtca tagagggcgg acctcttcct tttgctttcg atatcctgac
     1501 aacagcattc cattacggca accgggtatt tgtcgaatac ccagaagaaa tagtagacta
     1561 cttcaagcag tcgtttcctg agggttattc ttgggaacga agcatgagtt acgaagacgg
     1621 gggaatttgc ctcgccacaa acaatataac gatgaagaaa gacggcagca acacttttgt
     1681 caatgaaatt cgatttgatg gtacaaactt tcctgccaat ggtccagtta tgcagaggaa
     1741 gaccgtcaaa tgggagccat ccactgagaa aatgtatgtg cgtgatggag tgctgaaggg
     1801 tgatgttgaa atggctctgt tgcttgaagg aggtggccat taccgatgtg acttcagaac
     1861 tacttacaaa gcaaagaagg ttgtccagtt gccagactat cactacgtgg atcatcaaat
     1921 ggagataaca agccatgaca aggattacaa caaggttaag gcgtatgaac atgctaaggc
     1981 ttattccggg acctacaggg gcgccaagta tgaatttgaa gcctccggac tcagatctcg
     2041 agctgcggcc gccggatcca tggatgatga tatcgccgcg ctcgttgtcg acaacggctc
     2101 cggcatgtgc aaggccggct tcgcgggcga cgatgccccc cgggccgtct tcccctccat
     2161 cgtggggcgc cccaggcacc agggcgtgat ggtgggcatg ggtcagaagg attcctatgt
     2221 gggcgacgag gcccagagca agagaggcat cctcaccctg aagtacccca tcgagcacgg
     2281 catcgtcacc aactgggacg acatggagaa aatctggcac cacaccttct acaatgagct
     2341 gcgtgtggct cccgaggagc accccgtgct gctgaccgag gcccccctga accccaaggc
     2401 caaccgcgag aagatgaccc agatcatgtt tgagaccttc aacaccccag ccatgtacgt
     2461 tgctatccag gctgtgctat ccctgtacgc ctctggccgt accactggca tcgtgatgga
     2521 ctccggtgac ggggtcaccc acactgtgcc catctacgag gggtatgccc tcccccatgc
     2581 catcctgcgt ctggacctgg ctggccggga cctgactgac tacctcatga agatcctcac
     2641 cgagcgcggc tacagcttca ccaccacggc cgagcgggaa atcgtgcgtg acattaagga
     2701 gaagctgtgc tacgtcgccc tggacttcga gcaagagatg gccacggctg cttccagctc
     2761 ctccctggag aagagctacg agctgcctga cggccaggtc atcaccattg gcaatgagcg
     2821 gttccgctgc cctgaggcac tcttccagcc ttccttcctg ggcatggagt cctgtggcat
     2881 ccacgaaact accttcaact ccatcatgaa gtgtgacgtg gacatccgca aagacctgta
     2941 cgccaacaca gtgctgtctg gcggcaccac catgtaccct ggcattgccg acaggatgca
     3001 gaaggagatc actgccctgg cacccagcac aatgaagatc aagatcattg ctcctcctga
     3061 gcgcaagtac tccgtgtgga tcggcggctc catcctggcc tcgctgtcca ccttccagca
     3121 gatgtggatc agcaagcagg agtatgacga gtccggcccc tccatcgtcc accgcaaatg
     3181 cttctagtct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3241 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3301 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3361 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3421 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3481 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3541 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3601 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3661 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3721 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3781 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3841 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3901 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3961 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4021 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4081 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4141 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4201 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4261 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4321 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4381 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4441 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4501 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4561 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4621 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4681 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4741 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4801 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4861 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4921 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4981 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5041 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5101 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5161 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5221 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5281 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5341 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5401 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5461 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5521 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5581 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5641 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5701 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5761 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag at
//
