LOCUS       sequence-370036-        5496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a human siRNA-expressing plasmid targeting VAMP3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5496)
  TITLE     Zeng Yan Lab Plasmids 2022
REFERENCE   2  (bases 1 to 5496)
  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"
COMMENT     Sequence Label: pAAV.CMV.PI-si-VAMP3
FEATURES             Location/Qualifiers
     source          1..5496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    346..367
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(382..387,388..405,406..412)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    420..436
                     /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     425..447
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     444..460
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     444..460
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   498..627
                     /label=AAV2 ITR
     enhancer        768..1071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1072..1275
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1226..1246
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1454..1463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1460..1462,1463..1465,1466..2179)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1460..1462,1463..1465,1466..2176)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1505..1526)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1766..1785)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2113..2134
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2246..2266)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2664..2675)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2742..2759)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2748..2955
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3043..3172
                     /label=AAV2 ITR
     primer_bind     complement(3191..3208)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3191..3207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3200..3222)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3349..3804
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3436..3455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3646..3667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3830..3934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3935..4003,4004..4795)
                     /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(4153..4172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4966..5496,1..58)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5455..5474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
       61 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      121 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      181 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      241 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      301 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      361 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      421 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccagatttaa
      481 ttaaggcctt aattaggctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
      541 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
      601 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
      661 ctacgtagcc atgctctagg aagatcggaa ttcgccctta agctagctag ttattaatag
      721 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      781 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      841 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      901 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      961 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     1021 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1081 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     1141 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     1201 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     1261 tatataagca gagctggttt agtgaaccgt cagatcctgc agcatgggtt aacggatcca
     1321 tcgattctag aggtaccgct agcccgacag tcttcgatta ctgccagtgg tgaagccaca
     1381 gacactggca gtaatcgagg actgctgcct cgagtgacct tcagaccttg gcactggagg
     1441 tggcccggca gcggccgcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1501 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1561 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1621 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1681 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1741 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1801 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1861 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1921 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1981 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2041 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2101 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2161 ggacgagctg tacaagtaat aagcttggat ccaatcaacc tctggattac aaaatttgtg
     2221 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2281 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2341 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     2401 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2461 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2521 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2581 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     2641 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2701 tgctgccggc tctgcggcct cttccgcgtc ttcgagatct gcctcgactg tgccttctag
     2761 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2821 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2881 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2941 caggcatgct ggggactcga gttaagggcg aattcccgat aaggatcttc ctagagcatg
     3001 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     3061 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3121 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt
     3181 aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3241 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3301 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     3361 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3421 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3481 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3541 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3601 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3661 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3721 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3781 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4441 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5101 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5461 cgcctggtat ctttatagtc ctgtcgggtt tcgcca
//
