LOCUS       sequence-480245-        7499 bp    DNA     circular SYN 24-JUL-2025
DEFINITION  Expresses mBaoJin-tubulin fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7499)
  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 7499)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 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-Tubulin
FEATURES             Location/Qualifiers
     source          1..7499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..25)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      819..1407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1308..1327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1561..1578
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1695..1716
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1731..1736,1737..1754,1755..1761)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1769..1785
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1774..1796
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1793..1809
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1793..1809
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1858..1979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1869..1888)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1923..1942
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2153..2947)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2264..2283)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2874..2893
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3014..3343)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3028..3163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3082..3101)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3328..3348
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3357..3785)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3467..3488)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3679..3698
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3831..4055
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4044..4061
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        4081..4099
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4081..4098
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(5543..6244)
                     /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"
     regulatory      6241..6250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6269..6288)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6270..6288)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6333..6536)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6362..6382)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6537..6916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7088..7107
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7182..7286
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7287..7355,7356..7499,1..648)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
       61 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
      121 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
      181 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
      241 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
      301 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
      361 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
      421 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
      481 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
      541 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
      601 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
      661 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
      721 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
      781 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
      841 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      901 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      961 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1021 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1081 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1141 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1201 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1261 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1321 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1381 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1441 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     1501 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1561 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     1621 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     1681 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     1741 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     1801 agctatgacc atgattacgc caagctctag ctagaggtcg acggtataca gacatgataa
     1861 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1921 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg
     1981 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     2041 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     2101 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga
     2161 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     2221 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     2281 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     2341 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     2401 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     2461 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     2521 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     2581 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     2641 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     2701 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     2761 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     2821 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     2881 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     2941 caatcatgcg aaacgatcct catcctgtct cttgatcaga tccgaaaatg gatatacaag
     3001 ctcccgggag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     3061 atagctcaga ggcagaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     3121 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     3181 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     3241 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
     3301 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agaattaatt
     3361 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3421 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3481 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3541 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3601 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3661 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3721 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3781 cgcgtgggga taccccctag agccccagct ggttctttcc gcctcagaag ccatagagcc
     3841 caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
     3901 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
     3961 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
     4021 gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc gagctctagc
     4081 atttaggtga cactatagaa tagggccctc tagatccggt ggatccttag tattcctctc
     4141 cttcttcctc accctctcct tcaacagaat ccacaccaac ctcctcataa tccttctcaa
     4201 gggcagccat atcttcacgg gcctctgaaa actcgccttc ctccatcccc tcacccacgt
     4261 accagtgaac aaaggcacgc ttggcataca tcaggtcaaa cttgtggtcc aggcgagccc
     4321 aggcctcagc aatggctgtg gtgttgctca gcatgcacac agctctctgt accttggcca
     4381 ggtctccacc aggcaccaca gtgggaggct ggtagttgat gccaaccttg aagccagtgg
     4441 ggcaccaatc cacaaactgg atgctgcgct tggttttgat ggtggcaatg gcagcattga
     4501 catctttggg aaccacgtca ccacggtaca acaggcagca agccatgtat ttaccatggc
     4561 gagggtcaca tttcaccatc tggttggctg gctcaaagca agcattggtg atctctgcta
     4621 cagaaagctg ttcatggtag gctttctcag cagagatgac aggggcatat gtggccagag
     4681 ggaagtggat gcgggggtag ggcaccaggt tggtctggaa ttctgtcagg tcaacattca
     4741 gggctccatc aaatctcagg gaagcagtga tggaggacac aatctggcta ataaggcggt
     4801 taaggttagt gtaggttggg cgctcgatat cgaggtttct acgacagatg tcatagatgg
     4861 cctcattgtc taccatgaag gcacaatcag agtgctccag ggtggtgtgg gtggtgagga
     4921 tggagttgta gggctcaact acagctgtgg aaacctgggg tgctgggtaa atggagaact
     4981 ccagcttgga cttcttgcca taatcaactg agagacgttc catgagcagg gaggtgaacc
     5041 cagaaccagt tcccccacca aagctgtgga aaaccaagaa gccctgaaga ccggtgcact
     5101 ggtcagccag cttgcgaatt cggtccaaca caaggtcaat gatctccttg ccaatggtgt
     5161 agtgccctcg ggcatagtta ttggcagcat cttccttgcc tgtgatgagc tgctcagggt
     5221 ggaagagctg gcggtaggtg ccagtgcgaa cttcatcaat gactgtgggt tccaagtcta
     5281 caaacacagc ccggggcacg tgcttgccag cgcccgtctc actgaagaag gtgttgaagg
     5341 agtcatctcc tcccccaatg gtcttgtcac ttggcatctg gccatcgggc tggatgccgt
     5401 gttccaggca gtagagctcc cagcaggcat tgccaatctg gacaccagcc tggccaacgt
     5461 ggatggagat gcactcacgc actcgagatc ttcctcctga tccaccacct ccagaaccac
     5521 ctcctcctga tccaccacct cccttataca gctcgtccat gccctttaag tgtgcttcga
     5581 gagtctctga ttgacagata tgatcgcgtt cctcggcatc atctttgctt tgtgtgtatt
     5641 gtttacgaat ccagtgatat gggacatcag gtgctggttg attatgaaga ggcttgcaga
     5701 ttgtatattg gtgagcctcg acgtattttg atttatccga taataaagga taaagcaagg
     5761 tcactgggca ttctactcca tcatctctgg gtatttgtgg gacttcattc ggtaatgaga
     5821 cattcatatc tccagttaag actggactat tttctttaaa atcctgaccg gtaaattcta
     5881 caatgttgtg ataagtccca tttttcataa agtgttggtg ttttgcatgg atactcccat
     5941 cgcctttata ttgaatatga cgatcgtagg taaagcctcc gggcattact tcacgaaacc
     6001 agttcttcag tccactagga tatttggtat aatatttcat tccataacca aaggttgtcc
     6061 caagtgctgc ccatgacatc ggtagttttc cacttgtaca aacatatttt cctttatgag
     6121 aaccttcatg tgagttccct tcaccttcgc cttcaacggt aaacgatttg ccattgatgg
     6181 ttcctttaag ttgaaattta aatggtgtac tagccatgtt ttcctcctct ccctttgaga
     6241 ccatggtggc ggtaccaagc ttgggtctcc ctatagtgag tcgtattaat ttcgataagc
     6301 cagtaagcag tgggttctct agttagccag agagctctgc ttatatagac ctcccaccgt
     6361 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     6421 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     6481 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     6541 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     6601 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     6661 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     6721 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     6781 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     6841 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     6901 gtcaataatc aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc
     6961 ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa
     7021 attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt
     7081 aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc
     7141 ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     7201 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     7261 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     7321 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     7381 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     7441 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaa
//
