LOCUS       sequence-370200-        6147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a human siRNA-expressing plasmid targeting HBV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6147)
  TITLE     Zeng Yan Lab Plasmids 2022
REFERENCE   2  (bases 1 to 6147)
  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: pENN.AAV.SV40-si-HBV
FEATURES             Location/Qualifiers
     source          1..6147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     regulatory      1135..1144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1141..2571
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    2591..3179
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2644..2664)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3062..3073)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3222..3343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3233..3252)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3287..3306
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3505..3634
                     /label=AAV2 ITR
     rep_origin      complement(3871..4459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3951..3970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4630..5421,5422..5490))
                     /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     5253..5272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5491..5595)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5621..6076)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5758..5779)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5970..5989
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc gggggggggg gggggggggg gggggggggg
      601 gggggsgrgg sggrgrggkg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg gggygggrcm ggssgsksta gaggtaccgc tagcctcgag
     1021 gcgactgaga gtccaagagt cctcttgtga agccacagaa agaggactct tggattctta
     1081 gctgcctcga gtgaccttca gaccttggca ctggaggtgg cccggcagga tcccgccacc
     1141 atggtgagca agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag
     1201 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1261 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
     1321 atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc
     1381 cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1441 gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc
     1501 tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag
     1561 accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc
     1621 gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc
     1681 atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg
     1741 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc
     1801 cgccaccacc tgttcctggg gcatggcacc ggcagcaccg gcagcggcag ctccggcacc
     1861 gcctcctccg aggacaacaa catggccgtc atcaaagagt tcatgcgctt caaggtgcgc
     1921 atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1981 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
     2041 tgggacatcc tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc
     2101 gacatccccg attacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     2161 aacttcgagg acggcggtct ggtgaccgtg acccaggact cctccctgca ggacggcacg
     2221 ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc cgtaatgcag
     2281 aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg
     2341 aagggcgaga tccaccaggc cctgaagctg aaggacggcg gccgctacct ggtggagttc
     2401 aagaccatct acatggccaa gaagcccgtg caactgcccg gctactacta cgtggacacc
     2461 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgctcc
     2521 gagggccgcc accacctgtt cctgtacggc atggacgagc tgtacaagta agaattcgaa
     2581 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2641 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2701 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2761 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2821 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2881 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2941 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3001 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3061 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3121 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3181 tcgataccgt cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3241 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3301 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3361 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3421 ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc
     3481 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     3541 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3601 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc
     3661 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3721 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3781 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3841 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3901 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3961 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4021 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4081 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4141 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4201 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4261 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4321 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4381 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4441 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4501 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4561 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4621 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4801 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5281 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5461 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5521 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5581 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     5641 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     5701 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     5761 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     5821 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     5881 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5941 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     6001 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     6061 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     6121 gtgcgggcct cttcgctatt acgccag
//
