LOCUS       sequence-478764-        4791 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  Expresses P450-mBaoJin fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4791)
  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 4791)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 16, 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: pCytERM-mBaoJin-N1
FEATURES             Location/Qualifiers
     source          1..4791
                     /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      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             753..1454
                     /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    1579..1700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1616..1635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1670..1689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1707..2162)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1844..1865)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2056..2075
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2189..2293
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2295..2652
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2295..2486
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2318..2338)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2503..2638
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2687..3481
                     /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             2687..3481
                     /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(2741..2760)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3351..3370
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3669..3688)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3713..3760
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4089..4677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4578..4597
                     /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 ccggactcag atctatggac cctgtggtgg tgctggggct ctgtctctcc tgtttgcttc
      661 tcctttcact ctggaaacag agctatgggg gagggaagct tcgaattctg cagtcgacgg
      721 taccgcgggc ccgggatcca ccggtcgcca ccatggtctc aaagggagag gaggaaaaca
      781 tggctagtac accatttaaa tttcaactta aaggaaccat caatggcaaa tcgtttaccg
      841 ttgaaggcga aggtgaaggg aactcacatg aaggttctca taaaggaaaa tatgtttgta
      901 caagtggaaa actaccgatg tcatgggcag cacttgggac aacctttggt tatggaatga
      961 aatattatac caaatatcct agtggactga agaactggtt tcgtgaagta atgcccggag
     1021 gctttaccta cgatcgtcat attcaatata aaggcgatgg gagtatccat gcaaaacacc
     1081 aacactttat gaaaaatggg acttatcaca acattgtaga atttaccggt caggatttta
     1141 aagaaaatag tccagtctta actggagata tgaatgtctc attaccgaat gaagtcccac
     1201 aaatacccag agatgatgga gtagaatgcc cagtgacctt gctttatcct ttattatcgg
     1261 ataaatcaaa atacgtcgag gctcaccaat atacaatctg caagcctctt cataatcaac
     1321 cagcacctga tgtcccatat cactggattc gtaaacaata cacacaaagc aaagatgatg
     1381 ccgaggaacg cgatcatatc tgtcaatcag agactctcga agcacactta aagggcatgg
     1441 acgagctgta taagtaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     1501 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1561 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1621 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1681 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     1741 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1801 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1861 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     1921 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     1981 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2041 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2101 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2161 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2221 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2281 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2341 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2401 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2461 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2521 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2581 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2641 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2701 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2761 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2821 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     2881 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2941 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3001 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3061 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3121 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3181 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3241 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3301 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3361 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3421 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3481 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3541 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3601 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     3661 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3721 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3781 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3841 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3901 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     3961 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4021 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4081 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4141 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4201 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4261 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4321 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4381 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4441 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4501 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4561 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4621 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4681 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4741 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
