LOCUS       sequence-438234-        5365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector plasmid expressing mBaoJin under a short variant of the 
            CAG (sCAG) promoter. It also contains an optimised and truncated 
            version of WPRE as a regulatory element..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5365)
  TITLE     AAV fluorescent transgenes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5365)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: AAV-sCAG-mBaoJin-oPRE
FEATURES             Location/Qualifiers
     source          1..5365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        158..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        445..722
                     /label=chicken beta-actin promoter
     primer_bind     979..998
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1025..1049
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1050..1059
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1056..1757
                     /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"
     protein_bind    complement(1761..1785)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2396..2413)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2402..2609
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2617..2757
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2617..2746
                     /label=AAV2 ITR
     rep_origin      2832..3287
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2919..2938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3129..3150
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3304..3323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3423..3445
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3483..3501)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3569..3673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3674..3742,3743..4534)
                     /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     complement(3892..3911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4705..5293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5194..5213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5355..5365,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgcgt tacataactt acggtaaatg
      181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      481 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      541 cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg ggsggggggs
      601 ggggsggggs grggcggrga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      661 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      721 cgggagtcgc tgcgacgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      781 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      841 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      901 gaaagccttg aggggctccg ggagctagag cctctgctaa ccatgttcat gccttcttct
      961 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     1021 aattccaagt ttgtacaaaa aagcaggctg ccaccatggt ctcaaaggga gaggaggaaa
     1081 acatggctag tacaccattt aaatttcaac ttaaaggaac catcaatggc aaatcgttta
     1141 ccgttgaagg cgaaggtgaa gggaactcac atgaaggttc tcataaagga aaatatgttt
     1201 gtacaagtgg aaaactaccg atgtcatggg cagcacttgg gacaaccttt ggttatggaa
     1261 tgaaatatta taccaaatat cctagtggac tgaagaactg gtttcgtgaa gtaatgcccg
     1321 gaggctttac ctacgatcgt catattcaat ataaaggcga tgggagtatc catgcaaaac
     1381 accaacactt tatgaaaaat gggacttatc acaacattgt agaatttacc ggtcaggatt
     1441 ttaaagaaaa tagtccagtc ttaactggag atatgaatgt ctcattaccg aatgaagtcc
     1501 cacaaatacc cagagatgat ggagtagaat gcccagtgac cttgctttat cctttattat
     1561 cggataaatc aaaatacgtc gaggctcacc aatatacaat ctgcaagcct cttcataatc
     1621 aaccagcacc tgatgtccca tatcactgga ttcgtaaaca atacacacaa agcaaagatg
     1681 atgccgagga acgcgatcat atctgtcaat cagagactct cgaagcacac ttaaagggca
     1741 tggacgagct gtataagtag acccagcttt cttgtacaaa gtgggaattc gagcatctta
     1801 ccgccattta tacccatatt tgttctgttt ttcttgattt gggtatacat ttaaatgtta
     1861 ataaaacaaa atggtggggc aatcatttac atttttaggg atatgtaatt actagttcag
     1921 gtgtattgcc acaagacaaa catgttaaga aactttcccg ttatttacgc tctgttcctg
     1981 ttaatcaacc tctggattac aaaatttgtg aaagattgac tgatattctt aactatgttg
     2041 ctccttttac gctgtgtgga tatgctgctt tatagcctct gtatctagct attgcttccc
     2101 gtacggcttt cgttttctcc tccttgtata aatcctggtt gctgtctctt ttagaggagt
     2161 tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac gcaaccccca
     2221 ctggctgggg cattgccacc acctgtcaac tcctttctgg gactttcgct ttccccctcc
     2281 cgatcgccac ggcagaactc atcgccgcct gccttgcccg ctgctggaca ggggctaggt
     2341 tgctgggcac tgataattcc gtggtgttgt cgaattccta gagctcgctg atcagcctcg
     2401 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2461 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2521 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2581 tgggaagaga atagcaggca tgctggggag ggccgcagga acccctagtg atggagttgg
     2641 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2701 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     2761 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     2821 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2881 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     2941 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3001 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3061 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3121 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3181 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3241 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3301 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3361 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3421 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3481 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3541 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3601 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3661 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3721 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3781 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3841 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3901 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3961 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4021 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4081 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4141 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4201 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4261 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4321 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4381 tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4441 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4501 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4561 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4621 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4681 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4741 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4801 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4861 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4921 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4981 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5041 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5101 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5161 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5221 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5281 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5341 ttttgctcac atgtcctgca ggcag
//
