LOCUS       sequence-477130-        5537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EMTB-mBaoJin fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5537)
  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 5537)
  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"; doi: 
            "10.1038/s41592-024-02203-y"; journalName: "Nat Methods"; date: 
            "2024-04"; volume: "21"; issue: "4"; pages: "657-665"
COMMENT     Sequence Label: EMTB-mBaoJin
FEATURES             Location/Qualifiers
     source          1..5537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      271..280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             280..309
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1129..1138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1135..1836
                     /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    1961..2082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1998..2017)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2052..2071
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2089..2544)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2226..2247)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2438..2457
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2571..2675
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2677..3034
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2677..2868
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2700..2720)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2885..3020
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3069..3863
                     /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             3069..3863
                     /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(3123..3142)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3733..3752
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4051..4070)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4095..4142
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4471..5059
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4960..4979
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5234..5537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc gccaccatgg agcagaagct catctcagaa
      301 gaagacctcg gcgcgccagt gcgaagcgaa acagcacccg acagctacaa agtgcaagat
      361 aagaaaaatg cctccagccg ccctgcctct gcaatttcag gacaaaataa caaccactca
      421 ggaaataaac cagaccctcc gcctgtgtta cgtgttgatg accggcagcg gctggcccgg
      481 gagcgacgtg aggaacggga gaaacagcta gctgcaagag aaatagtgtg gttagaaaga
      541 gaagagcgag ccaggcagca ctacgagaag cacctggaag agcggaagaa gaggttggag
      601 gagcagaggc agaaggagga gcggaggagg gctgctgtgg aggagaagcg gaggcagaga
      661 cttgaggagg acaaagaacg ccacgaagct gttgtacggc gcacaatgga aaggagccag
      721 aagccaaaac agaagcataa ccgttggtcg tggggaggct ctctccatgg gagccctagc
      781 atccacagtg cagatccaga caggcggtca gtttccacca tgaatctttc gaaatatgtt
      841 gatcccgtca ttagcaagcg gctctcctct tcatctgcaa ctttactaaa ttctccagat
      901 agagctcgcc gcctgcagct cagcccatgg gagagcagcg ttgttaacag actcctgacg
      961 cccacacatt cgttcctggc cagaagtaaa agcacagctg ccttgtctgg agaagcagca
     1021 tcttgcagcc ccatcatcat gccctacaaa gctgcacact ctagaaattc gatggatcga
     1081 ccaaaactct ttgtaacacc acctgagggc tcttcggatc caccggtcgc caccatggtc
     1141 tcaaagggag aggaggaaaa catggctagt acaccattta aatttcaact taaaggaacc
     1201 atcaatggca aatcgtttac cgttgaaggc gaaggtgaag ggaactcaca tgaaggttct
     1261 cataaaggaa aatatgtttg tacaagtgga aaactaccga tgtcatgggc agcacttggg
     1321 acaacctttg gttatggaat gaaatattat accaaatatc ctagtggact gaagaactgg
     1381 tttcgtgaag taatgcccgg aggctttacc tacgatcgtc atattcaata taaaggcgat
     1441 gggagtatcc atgcaaaaca ccaacacttt atgaaaaatg ggacttatca caacattgta
     1501 gaatttaccg gtcaggattt taaagaaaat agtccagtct taactggaga tatgaatgtc
     1561 tcattaccga atgaagtccc acaaataccc agagatgatg gagtagaatg cccagtgacc
     1621 ttgctttatc ctttattatc ggataaatca aaatacgtcg aggctcacca atatacaatc
     1681 tgcaagcctc ttcataatca accagcacct gatgtcccat atcactggat tcgtaaacaa
     1741 tacacacaaa gcaaagatga tgccgaggaa cgcgatcata tctgtcaatc agagactctc
     1801 gaagcacact taaagggcat ggacgagctg tataagtagg cggccgcgac tctagatcat
     1861 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1921 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1981 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2041 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2101 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2161 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2221 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2281 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2341 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2401 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2461 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2521 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2581 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2641 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2701 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2761 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2881 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2941 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3001 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3061 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3121 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3181 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3241 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3301 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3361 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3421 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3481 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3541 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3601 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3661 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3721 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3781 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3841 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3901 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3961 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4021 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4081 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4141 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4201 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4261 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4321 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4381 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4441 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4501 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4561 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4621 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4681 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4741 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4801 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4861 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4921 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4981 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5041 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5101 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5161 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     5221 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5281 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5341 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5401 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5461 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5521 tcatcgctat taccatg
//
