LOCUS       sequence-480261-        5716 bp    DNA     circular SYN 24-JUL-2025
DEFINITION  Expresses mBaoJin and FusionRed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  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 5716)
  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: pAAV-CAG-mBaoJin-P2A-FusionRed
FEATURES             Location/Qualifiers
     source          1..5716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     482..504
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     566..585
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      624..633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             630..1331
                     /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"
     CDS             1356..1412
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1425..2123
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     misc_feature    2148..2736
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2201..2221)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2619..2630)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2779..2900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2844..2863
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3279..3867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3359..3378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4038..4829,4830..4898))
                     /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"
     primer_bind     4661..4680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4899..5003)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5029..5484)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5166..5187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5378..5397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5556..5685
                     /label=AAV2 ITR
ORIGIN
        1 atctacgtag ccatgctcta ggaagagtac cattgacgtc aataatgacg tatgttccca
       61 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      121 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      181 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      241 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      301 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      361 aattattttg tgcagcgatg ggggcggggg gggggggggg ggggggccgg gggggggggg
      421 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggcagggcg
      481 gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc
      541 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
      601 ggcaaagaat tggatccggt accgccacca tggtctcaaa gggagaggag gaaaacatgg
      661 ctagtacacc atttaaattt caacttaaag gaaccatcaa tggcaaatcg tttaccgttg
      721 aaggcgaagg tgaagggaac tcacatgaag gttctcataa aggaaaatat gtttgtacaa
      781 gtggaaaact accgatgtca tgggcagcac ttgggacaac ctttggttat ggaatgaaat
      841 attataccaa atatcctagt ggactgaaga actggtttcg tgaagtaatg cccggaggct
      901 ttacctacga tcgtcatatt caatataaag gcgatgggag tatccatgca aaacaccaac
      961 actttatgaa aaatgggact tatcacaaca ttgtagaatt taccggtcag gattttaaag
     1021 aaaatagtcc agtcttaact ggagatatga atgtctcatt accgaatgaa gtcccacaaa
     1081 tacccagaga tgatggagta gaatgcccag tgaccttgct ttatccttta ttatcggata
     1141 aatcaaaata cgtcgaggct caccaatata caatctgcaa gcctcttcat aatcaaccag
     1201 cacctgatgt cccatatcac tggattcgta aacaatacac acaaagcaaa gatgatgccg
     1261 aggaacgcga tcatatctgt caatcagaga ctctcgaagc acacttaaag ggcatggacg
     1321 agctgtataa gcgaccggtt cacgtgggaa gtggggctac aaacttctca ctgctgaaac
     1381 aggccgggga tgtggaggaa aacccaggac cacacgtgac tagtatggtg agcgagctga
     1441 ttaaggagaa catgcccatg aagctgtaca tggagggcac cgtgaacaac caccacttca
     1501 agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg
     1561 tcgtcgaggg cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg
     1621 gcagcagaac cttcatcaag caccctccgg gcatccccga cttctttaag cagtccttcc
     1681 ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta
     1741 cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaaggtt agaggggtga
     1801 acttcccagc caacggccct gtgatgcaga agaaaacact cggctgggag gcctccaccg
     1861 agacgatgta ccctgctgac ggaggactgg aaggcgcatg tgacatggct ctgaagctcg
     1921 tgggaggagg acacctgatc tgcaaccttg agacaacata cagatccaag aaacccgcta
     1981 cgaacctcaa gatgcccggc gtctacaacg tggaccacag actggaaaga atcaaggagg
     2041 ccgacgatga gacatacgtc gagcagcacg aggtggctgt ggccagatac tctactggtg
     2101 gcgctggtga tggaggtaaa taagaattcg atatcaagct tatcgataat caacctctgg
     2161 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2221 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2281 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     2341 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     2401 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     2461 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2521 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     2581 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     2641 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     2701 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc
     2761 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2821 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2881 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2941 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     3001 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     3061 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgac
     3121 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3181 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4321 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4921 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4981 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     5041 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     5101 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     5161 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     5221 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     5281 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     5341 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     5401 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     5461 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     5521 ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg ctcactgagg
     5581 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     5641 gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta
     5701 acccgccatg ctactt
//
