LOCUS       sequence-370035-        5502 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 5502)
  TITLE     Zeng Yan Lab Plasmids 2022
REFERENCE   2  (bases 1 to 5502)
  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-HBV
FEATURES             Location/Qualifiers
     source          1..5502
                     /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      1460..1469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1466..1468,1469..1471,1472..2185)
                     /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(1466..1468,1469..1471,1472..2182)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1511..1532)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1772..1791)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2119..2140
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2252..2272)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2670..2681)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2748..2765)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2754..2961
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3049..3178
                     /label=AAV2 ITR
     primer_bind     complement(3197..3214)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3197..3213)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3206..3228)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3355..3810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3442..3461)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3652..3673
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3836..3940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3941..4009,4010..4801)
                     /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(4159..4178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4972..5502,1..58)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5461..5480
                     /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 agcctcgagg cgactgagag tccaagagtc ctcttgtgaa
     1381 gccacagaaa gaggactctt ggattcttag ctgcctcgag tgaccttcag accttggcac
     1441 tggaggtggc ccggcagcgg ccgccatggt gagcaagggc gaggagctgt tcaccggggt
     1501 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1561 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1621 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1681 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1741 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1801 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1861 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1921 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     1981 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2041 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     2101 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2161 cggcatggac gagctgtaca agtaataagc ttggatccaa tcaacctctg gattacaaaa
     2221 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2281 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2341 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2401 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2461 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2521 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2581 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     2641 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2701 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg agatctgcct cgactgtgcc
     2761 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2821 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2881 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2941 caatagcagg catgctgggg actcgagtta agggcgaatt cccgataagg atcttcctag
     3001 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     3061 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3121 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc
     3181 ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3241 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3301 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     3361 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3421 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3481 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3541 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3601 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3661 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3721 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3781 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     3841 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3901 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3961 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4021 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4081 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4141 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4201 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4261 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4321 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4381 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4441 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4501 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4561 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4621 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4681 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4741 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4801 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4861 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4921 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4981 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5041 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5101 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5161 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5221 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5281 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5341 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5401 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5461 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc ca
//
