LOCUS       sequence-480517-        5763 bp    DNA     circular SYN 25-JUL-2025
DEFINITION  Expresses mBaoJin-actin fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5763)
  AUTHORS   Zhang H, Lesnov GD, Subach OM, Zhang W, Kuzmicheva TP, Vlaskina AV, 
            Samygina VR, Chen L, Ye X, Nikolaeva AY, Gabdulkhakov A, Papadaki S,
            Qin W, Borshchevskiy V, Perfilov MM, Gavrikov AS, Drobizhev M, 
            Mishin AS, Piatkevich KD, Subach FV
  TITLE     Bright and stable monomeric green fluorescent protein derived from 
            StayGold.
  JOURNAL   Nat Methods. 2024 Apr;21(4):657-665. doi: 
            10.1038/s41592-024-02203-y. Epub 2024 Feb 26.
  PUBMED    38409224
REFERENCE   2  (bases 1 to 5763)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-024-02203-y"; journalName: "Nat Methods"; date: 
            "2024-04"; volume: "21"; issue: "4"; pages: "657-665"
COMMENT     Sequence Label: pmBaoJin-Actin
FEATURES             Location/Qualifiers
     source          1..5763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1314
                     /codon_start=1
                     /product="bright and photostable monomeric green 
                     fluorescent protein derived from StayGold (Zhang et al., 
                     2024)"
                     /label=mBaoJin
                     /label=mStayGold
                     /note="/vntifkey=21"
                     /translation="MVSKGEEENMASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTTFGYGMKYYTKYPSGLKNWFREVMPGGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVPQIPRDDGVECPVTL
                     LYPLLSDKSKYVEAHQYTICKPLHNQPAPDVPYHWIRKQYTQSKDDAEERDHICQSETL
                     EAHLKGMDELYK"
     polyA_signal    2623..2744
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2660..2679)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2714..2733
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2751..3206)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2888..2909)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3100..3119
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3233..3337
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3362..3382)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3406..3616
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3428..3624
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3475..3610
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3659..4453
                     /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             3659..4453
                     /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(3713..3732)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4323..4342
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4641..4660)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4685..4732
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5061..5649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5550..5569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtctc aaagggagag gaggaaaaca tggctagtac accatttaaa
      661 tttcaactta aaggaaccat caatggcaaa tcgtttaccg ttgaaggcga aggtgaaggg
      721 aactcacatg aaggttctca taaaggaaaa tatgtttgta caagtggaaa actaccgatg
      781 tcatgggcag cacttgggac aacctttggt tatggaatga aatattatac caaatatcct
      841 agtggactga agaactggtt tcgtgaagta atgcccggag gctttaccta cgatcgtcat
      901 attcaatata aaggcgatgg gagtatccat gcaaaacacc aacactttat gaaaaatggg
      961 acttatcaca acattgtaga atttaccggt caggatttta aagaaaatag tccagtctta
     1021 actggagata tgaatgtctc attaccgaat gaagtcccac aaatacccag agatgatgga
     1081 gtagaatgcc cagtgacctt gctttatcct ttattatcgg ataaatcaaa atacgtcgag
     1141 gctcaccaat atacaatctg caagcctctt cataatcaac cagcacctga tgtcccatat
     1201 cactggattc gtaaacaata cacacaaagc aaagatgatg ccgaggaacg cgatcatatc
     1261 tgtcaatcag agactctcga agcacactta aagggcatgg acgagctgta taagtccgga
     1321 ctcagatctg gcagcggtgg aggcagcgca tccggcggaa gcggaagcga tgatgatatc
     1381 gccgcgctcg tcgtcgacaa cggctccggc atgtgcaagg ccggcttcgc gggcgacgat
     1441 gccccccggg ccgtcttccc ctccatcgtg gggcgcccca ggcaccaggg cgtgatggtg
     1501 ggcatgggtc agaaggattc ctatgtgggc gacgaggccc agagcaagag aggcatcctc
     1561 accctgaagt accccatcga gcacggcatc gtcaccaact gggacgacat ggagaaaatc
     1621 tggcaccaca ccttctacaa tgagctgcgt gtggctcccg aggagcaccc cgtgctgctg
     1681 accgaggccc ccctgaaccc caaggccaac cgcgagaaga tgacccagat catgtttgag
     1741 accttcaaca ccccagccat gtacgttgct atccaggctg tgctatccct gtacgcctct
     1801 ggccgtacca ctggcatcgt gatggactcc ggtgacgggg tcacccacac tgtgcccatc
     1861 tacgaggggt atgccctccc ccatgccatc ctgcgtctgg acctggctgg ccgggacctg
     1921 actgactacc tcatgaagat cctcaccgag cgcggctaca gcttcaccac cacggccgag
     1981 cgggaaatcg tgcgtgacat taaggagaag ctgtgctacg tcgccctgga cttcgagcaa
     2041 gagatggcca cggctgcttc cagctcctcc ctggagaaga gctacgagct gcctgacggc
     2101 caggtcatca ccattggcaa tgagcggttc cgctgccctg aggcactctt ccagccttcc
     2161 ttcctgggca tggagtcctg tggcatccac gaaactacct tcaactccat catgaagtgt
     2221 gacgtggaca tccgcaaaga cctgtacgcc aacacagtgc tgtctggcgg caccaccatg
     2281 taccctggca ttgccgacag gatgcagaag gagatcactg ccctggcacc cagcacaatg
     2341 aagatcaaga tcattgctcc tcctgagcgc aagtactccg tgtggatcgg cggctccatc
     2401 ctggcctcgc tgtccacctt ccagcagatg tggatcagca agcaggagta tgacgagtcc
     2461 ggcccctcca tcgtccaccg caaatgcttc tagggatcca ccggatctag ataactgatc
     2521 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2581 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2641 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2701 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     2761 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2821 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2881 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2941 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3001 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3061 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3121 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3181 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3241 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3301 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3361 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3421 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3481 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3541 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3601 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     3661 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3721 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3781 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3841 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3901 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3961 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4021 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4081 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4141 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4201 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4261 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4321 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4381 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4441 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4501 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4561 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4621 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4681 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     4741 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4801 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4861 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     4921 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4981 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5041 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5101 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5161 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5221 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5281 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5341 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5401 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5461 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5521 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5581 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5641 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5701 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     5761 cat
//
