LOCUS       sequence-477195-        5073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mBaoJin in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5073)
  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 5073)
  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: pBADHisB-Sumo-mBaoJin
FEATURES             Location/Qualifiers
     source          1..5073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             424..717
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             724..1425
                     /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"
     primer_bind     complement(1474..1491)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1644..1730
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1822..1849
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1868..1959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1960..2028,2029..2820)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2178..2197)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2991..3579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3480..3499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3733..3750
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3765..3905
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3831..3850)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3991..4013
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4169..5047)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgatgtcgg actcagaagt caatcaagaa gctaagccag aggtcaagcc agaagtcaag
      481 cctgagactc acatcaattt aaaggtgtcc gatggatctt cagagatttt cttcaagatc
      541 aaaaagacca ctcctttaag aaggctgatg gaagcgttcg ctaaaagaca gggtaaggaa
      601 atggactcct taagattctt gtacgacggt attagaatcc aagctgatca gacccctgaa
      661 gatttggaca tggaggataa cgatattatt gaggctcaca gagaacagat tggtggaaga
      721 tctatggtct caaagggaga ggaggaaaac atggctagta caccatttaa atttcaactt
      781 aaaggaacca tcaatggcaa atcgtttacc gttgaaggcg aaggtgaagg gaactcacat
      841 gaaggttctc ataaaggaaa atatgtttgt acaagtggaa aactaccgat gtcatgggca
      901 gcacttggga caacctttgg ttatggaatg aaatattata ccaaatatcc tagtggactg
      961 aagaactggt ttcgtgaagt aatgcccgga ggctttacct acgatcgtca tattcaatat
     1021 aaaggcgatg ggagtatcca tgcaaaacac caacacttta tgaaaaatgg gacttatcac
     1081 aacattgtag aatttaccgg tcaggatttt aaagaaaata gtccagtctt aactggagat
     1141 atgaatgtct cattaccgaa tgaagtccca caaataccca gagatgatgg agtagaatgc
     1201 ccagtgacct tgctttatcc tttattatcg gataaatcaa aatacgtcga ggctcaccaa
     1261 tatacaatct gcaagcctct tcataatcaa ccagcacctg atgtcccata tcactggatt
     1321 cgtaaacaat acacacaaag caaagatgat gccgaggaac gcgatcatat ctgtcaatca
     1381 gagactctcg aagcacactt aaagggcatg gacgagctgt ataagtagga attcgaagct
     1441 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     1501 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     1561 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     1621 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1681 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     1741 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     1801 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     1861 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1921 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1981 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2041 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2101 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2161 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2221 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2281 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2341 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2401 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2461 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2521 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2581 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2641 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2701 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2761 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2821 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2881 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2941 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3001 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3061 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3121 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3181 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3241 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3301 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3361 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3421 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3481 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3541 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3601 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3661 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3721 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3781 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     3841 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3901 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3961 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4021 cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg
     4081 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
     4141 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
     4201 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     4261 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     4321 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     4381 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     4441 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     4501 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     4561 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     4621 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     4681 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     4741 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     4801 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     4861 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     4921 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     4981 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     5041 cagccatact tttcatactc ccgccattca gag
//
