LOCUS       sequence-139082-        5859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5859)
  AUTHORS   Geng X, Huang C, Qin Y, McCombs JE, Yuan Q, Harry BL, Palmer AE, Xia
            NS, Xue D
  TITLE     Hepatitis B virus X protein targets Bcl-2 proteins to increase 
            intracellular calcium, required for virus replication and cell death
            induction.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Nov 6;109(45):18471-6. doi: 
            10.1073/pnas.1204668109. Epub 2012 Oct 22.
  PUBMED    23091012
REFERENCE   2  (bases 1 to 5859)
  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"; doi: 
            "10.1073/pnas.1204668109"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2012-11-6- 6"; volume: "109"; issue: "45"; pages: "18471-6"
COMMENT     Sequence Label: pcDNA3.1-Flag HBx
FEATURES             Location/Qualifiers
     source          1..5859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1453..1470)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1459..1683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1729..2157
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1816..1835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2026..2047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2166..2186)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2171..2500
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2351..2486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2413..2432
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2567..3361
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2621..2640)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3231..3250
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3535..3656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3572..3591)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3626..3645
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3705..3721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3705..3721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3718..3740)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3729..3745
                     /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)."
     promoter        complement(join(3753..3759,3760..3777,3778..3783))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3798..3819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3936..3953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4107..4692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4187..4206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4863..5654,5655..5723))
                     /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     5486..5505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5724..5828)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atggactaca aggatgacga tgacaagatg gctgctaggc tgtactgcca actggatcct
      961 tcgcgggacg tcctttgttt acgtcccgtc ggcgctgaat cccgcggacg acccctctcg
     1021 gggccgcttg ggactctctc gtccccttct ccgtctgccg ttccagccga ccacggggcg
     1081 cacctctctt tacgcggtct ccccgtctgt gccttctcat ctgccggtcc gtgtgcactt
     1141 cgcttcacct ctgcacgttg catggagacc accgtgaacg cccatcagat cctgcccaag
     1201 gtcttacata agaggactct tggactccca gcaatgtcaa cgaccgacct tgaggcctac
     1261 ttcaaggact gtgtgtttaa ggactgggag gagctggggg aggagattag gttaaaggtc
     1321 tttgtattag gaggctgtag gcataaattg gtctgcgcac cagcaccatg caactttttc
     1381 acctctgcct aagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa
     1441 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1501 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1561 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1621 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     1681 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     1741 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1801 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1861 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1921 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1981 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2041 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2101 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2161 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2221 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2281 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2341 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2401 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2461 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     2521 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2581 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2641 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2701 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     2761 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2821 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2881 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2941 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3001 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3061 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3121 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3181 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3241 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3301 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3361 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3421 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3481 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3661 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3721 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3781 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3841 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3901 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3961 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4021 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4081 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4141 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4201 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4261 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4321 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4381 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4441 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4501 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4561 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4621 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5461 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5701 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5761 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5821 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
