LOCUS       sequence-111012-        6763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses HBx in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6763)
  AUTHORS   Cha MY, Kim CM, Park YM, Ryu WS
  TITLE     Hepatitis B virus X protein is essential for the activation of 
            Wnt/beta-catenin signaling in hepatoma cells.
  JOURNAL   Hepatology. 2004 Jun;39(6):1683-93.
  PUBMED    15185310
REFERENCE   2  (bases 1 to 6763)
  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: 
            "Hepatology."; date: "2004-06"; volume: "39(6)"; pages: "1683-93"
COMMENT     Sequence Label: HBx
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             481..546
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    557..1179
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(756..775)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1203..1223)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1221..1518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             1572..2366
                     /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(1626..1645)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2236..2255
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3018..3152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3055..3074)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3109..3128
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3157..3176)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3158..3176)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3190..3206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3190..3206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3203..3225)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3214..3230
                     /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(3238..3244,3245..3262,3263..3268))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3283..3304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3421..3438)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3592..4180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3672..3691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4351..5142,5143..5211))
                     /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     4974..4993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5212..5316)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5343..5798
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5430..5449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5640..5661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5924..5946
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5938..5955
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5939..5955
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6116..6495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6496..6699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6650..6670
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggctgcta ggctgtgctg ccaactggat cctgcgcggg acgtcctttg tttacgtccc
       61 gtcggcgctg aatcctgcgg acgacccttc tcggggtcgc ttgggactct ctcgtcccct
      121 tctccgtctg ccgttccgac cgaccacggg gcgcacctct ctttacgcgg actccccgtc
      181 tgtgccttct catctgccgg accgtgtgca cttcgcttca cctctgcacg tcgcatggag
      241 accaccgtga acgcccacca aatattgccc aaggtcttac ataagaggac tcttggactc
      301 tcagcaatgt caacgaccga ccttgaggca tacttcaaag actgtttgtt taaagactgg
      361 gaggagttgg gggaggagat taggttaaag gtctttgtac taggaggctg taggcataaa
      421 ttggtctgcg caccagcacc atgcaacttt ttcacctctg cctctagagg atcccgggct
      481 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
      541 gacaagtagt gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
      601 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
      661 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
      721 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
      781 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
      841 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
      901 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
      961 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     1021 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     1081 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     1141 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
     1201 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1261 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     1321 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1381 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1441 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     1501 gtgaggaggc ttttttggag gaattgatca gcttgggatc tgatcaagag acaggatgag
     1561 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     1621 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     1681 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     1741 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     1801 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     1861 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     1921 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     1981 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     2041 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     2101 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     2161 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     2221 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     2281 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     2341 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     2401 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     2461 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     2521 ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt tggttcagct gctgcctgag
     2581 gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag
     2641 cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca cgatggccgc
     2701 tttggtcgac ccggacggga cgctcctgcg cctgatacag aacgaattgc ttgcaggcat
     2761 ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg cgtggatgga gcgctggcgc
     2821 ctgctgcgcg acggcgagct gctcaccacc cactcgccaa gctggaaccg taaaaaggcc
     2881 gcgttgctgg cgtttttcca taggctccgc cgatcataat cagccatacc acatttgtag
     2941 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3001 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3061 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3121 aactcatcaa tgtatcttat catgtctgga tcaattccct atagtgagtc gtattaaatt
     3181 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3241 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3301 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3361 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3421 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3481 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3541 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3601 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3661 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3721 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3781 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3841 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3901 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3961 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4021 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4081 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4141 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4201 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4261 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4321 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4381 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4441 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4501 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4561 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4621 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4681 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4741 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4801 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4861 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4921 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4981 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5041 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5101 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5161 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5221 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5281 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5341 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5401 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5461 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5521 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5581 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5641 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5701 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5761 taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc
     5821 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     5881 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     5941 aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca attagccata
     6001 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     6061 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat
     6121 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     6181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6361 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6421 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     6481 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
     6541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     6601 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
     6661 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagaat
     6721 taagcttctc tgttgtccta tcccgcaaat atacatcgtt tcc
//
