LOCUS       sequence-478810-        7494 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  Expresses mBaoJin-NUP98 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7494)
  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 7494)
  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: pmBaoJin-NUP98
FEATURES             Location/Qualifiers
     source          1..7494
                     /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    4282..4403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4319..4338)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4373..4392
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4410..4865)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4547..4568)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4759..4778
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4892..4996
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4998..5355
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4998..5189
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5021..5041)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5206..5341
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5390..6184
                     /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             5390..6184
                     /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(5444..5463)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6054..6073
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6372..6391)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6416..6463
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6792..7380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7281..7300
                     /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 caagtccggc
     1321 cggactcaga tctcgagcat gtttaacaaa tcatttggaa caccctttgg gggtggcaca
     1381 ggtggctttg gcacaacttc aacatttgga cagaatactg gctttggcac tactagtgga
     1441 ggggcatttg gaacatctgc atttggttct agcaacaata ctggaggcct ctttggaaat
     1501 tcacagacta aaccaggagg attgtttgga accagttcat ttagccagcc agctacctcc
     1561 acaagcactg gctttgggtt tggtacgtca acaggaacag caaatacctt gtttggaact
     1621 gcaagcacag ggaccagtct cttctcatcc caaaacaatg cctttgcaca aaataaacca
     1681 actggctttg gcaattttgg aaccagtact agcagtggag gactctttgg aaccacaaat
     1741 accacctcta atccttttgg cagcacatct ggctccctct ttgggccaag tagttttaca
     1801 gctgctccta ctgggactac tattaaattt aaccctccaa ctggtacaga tactatggtc
     1861 aaagctggag ttagcactaa cataagtacc aagcaccagt gtattactgc tatgaaagaa
     1921 tatgaaagca agtcactaga ggaacttcgt ttagaggatt atcaggctaa caggaagggc
     1981 ccacagaacc aggtgggagc aggtaccaca actggcttgt ttgggtcttc tccagccact
     2041 tccagcgcaa caggactctt cagctcctcc accactaatt caggctttgc atatggtcag
     2101 aacaaaactg cctttggaac tagtacaact ggatttggaa caaatccagg tggtctcttt
     2161 ggccaacaga atcagcagac taccagcctc ttcagcaaac catttggcca ggctacaacc
     2221 acccagaaca ctggcttttc ctttggtaat accagcacca taggacagcc aagcaccaac
     2281 accatgggat tatttggagt aacccaagcc tcacagcctg gaggtctttt tgggacagct
     2341 acaaacacca gcactgggac agcatttgga acaggaacag gtctctttgg gcagaccaat
     2401 actggatttg gtgctgttgg ttcgaccctg tttggcaata acaagcttac tacatttgga
     2461 agcagcacaa ccagtgcacc ttcatttggt acaaccagtg gcgggctctt tggtaacaaa
     2521 ccaaccctga ctttaggaac caatacaaac acttctaatt ttggttttgg cacaaatacc
     2581 agtgggaata gtatttttgg aagtaaacca gcacctggga ctcttggaac tgggcttggt
     2641 gcaggatttg gaacagctct tggtgctgga caggcatctt tgtttgggaa caaccaacct
     2701 aagattggag ggcctcttgg tacaggagcc tttggggccc ctggatttaa tactacgaca
     2761 gccactttgg gctttggagc cccccaggcc ccagtagctt tgacagatcc aaatgcttct
     2821 gctgcccagc aggctgttct ccagcagcac atcaatagtc taacatactc accttttgga
     2881 gactctcctc tcttccggaa tccgatgtca gaccctaaga agaaggaaga gagattgaaa
     2941 ccaacaaatc cagcagccca gaaggctctt actacaccta ctcattataa actgacaccc
     3001 cgccctgcca ctagagtccg gccaaaggct ttacaaacaa caggcacagc caagtcacat
     3061 ctctttgatg ggctggatga cgatgaacca tccctagcca atggagcatt catgcccaag
     3121 aagagcatta agaagttggt tttgaagaac cttaataata gcaatctctt ttctcctgtt
     3181 aatcgtgatt cagaaaatct agcttcacca tctgaatatc cagaaaatgg agagagattt
     3241 agtttcctaa gcaaacctgt tgatgagaat caccagcagg atggagatga agattccctt
     3301 gtttcacatt tttatactaa ccctattgcc aaacctattc ctcaaacccc agaaagtgct
     3361 ggaaataaac acagcaacag caacagtgtg gatgatacca ttgttgcatt aaacatgcgt
     3421 gctgctttgc gaaatgggct ggaaggaagc agtgaagaaa cgtcttttca tgatgagtca
     3481 cttcaggatg accgagaaga aatagaaaat aattcttacc atatgcaccc agcaggtatt
     3541 attctcacta aggttggtta ctatactatt ccatctatgg atgaccttgc taaaattacc
     3601 aatgaaaaag gagagtgcat tgtctctgat ttcactattg gtcggaaagg ttatggttca
     3661 atctattttg aaggagatgt gaatttgaca aatctaaatt tggatgatat tgtgcatatc
     3721 cggaggaaag aagtagttgt ctacttagat gataaccaaa aaccacctgt gggtgaaggg
     3781 ctaaatagga aggctgaagt tacattggat ggagtttggc caacagataa aacatctcgt
     3841 tgtttaataa agagcccaga tcgccttgct gatatcaact atgaaggaag attggaagca
     3901 gtttcaagga aacagggagc tcaattcaaa gaataccggc ctgaaactgg ttcttgggtg
     3961 tttaaggtct cccatttttc taagtatggc cttcaggatt ctgatgaaga ggaggaggag
     4021 catccgtcta aaactagtac aaagaagttg aagactgctc ctttgcctcc tgcaagccag
     4081 actacgccct tgcagatggc tcttaatggc aaacctgcac ctccacctca ggtagagaaa
     4141 aaaggacagt gaggatccac cggatctaga taactgatca taatcagcca taccacattt
     4201 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4261 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4321 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4381 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     4441 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4501 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4561 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4621 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4681 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4741 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4801 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4861 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4921 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4981 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     5041 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5101 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5161 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5221 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5281 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5341 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5401 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5461 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5521 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     5581 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5641 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5701 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5761 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5821 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5881 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5941 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6001 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6061 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6121 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6181 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6241 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6301 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6361 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     6421 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     6481 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     6541 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     6601 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     6661 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6721 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6781 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6841 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6901 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6961 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7021 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7081 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7141 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7201 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7261 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7321 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7381 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7441 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
