LOCUS       Exported                6607 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses HBV Large envelope glycoprotein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    L-HBsAg
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6607)
  AUTHORS   Wang H, Ryu WS
  TITLE     Hepatitis B virus polymerase blocks pattern recognition receptor 
            signaling via interaction with DDX3: implications for immune 
            evasion.
  JOURNAL   PLoS Pathog. 2010 Jul 15;6(7):e1000986. doi: 
            10.1371/journal.ppat.1000986.
  PUBMED    20657822
REFERENCE   2  (bases 1 to 6607)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6607
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1280..2125
                     /codon_start=1
                     /product="middle (pre-S2/S) envelope protein of hepatitis B
                     virus"
                     /label=HBsAg (pre-S2/S)
                     /note="hepatitis B surface antigen"
                     /translation="MQWNSTTFHQTLQDPRVRGLYFPAGGSSSGTVNPVLTTASPLSSI
                     FSRIGDPALNMENITSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGTTVCL
                     GQNSQSPTSNHSPTSCPPTCPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGMLPVCP
                     LIPGSSTTSTGPCRTCMTTAQGTSMYPSCCCTKPSDGNCTCIPIPSSWAFGKFLWEWAS
                     ARFSWLSLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYSILSPFLPLLPIFFCLWVYI
                     "
     promoter        complement(2160..2178)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2161..2178)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2198..2215)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2204..2428
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2474..2902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2561..2580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2771..2792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2911..2931)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2916..3245
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3096..3231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3158..3177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3312..4106
                     /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(3366..3385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3976..3995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4280..4401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4317..4336)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4371..4390
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4450..4466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4450..4466)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4463..4485)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4474..4490
                     /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(4498..4528)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4543..4564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4681..4698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4852..5440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4932..4951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5611..6471)
                     /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     6234..6253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6472..6576)
                     /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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atcccatggg
      961 gcagaatctt tccaccagca atcctctggg attctttccc gaccaccagt tggatccagc
     1021 cttcagagca aacaccgcaa atccagattg ggacttcaat cccaacaagg acacctggcc
     1081 agacgccaac aaggtaggag ctggagcatt cgggctgggt ttcaccccac cgcacggagg
     1141 ccttttgggg tggagccctc aggctcaggg catactacaa actttgccag caaatccgcc
     1201 tcctgcctcc accaatcgcc agtcaggaag gcagcctacc ccgctgtctc cacctttgag
     1261 aaacactcat cctcaggcca tgcagtggaa ttccacaacc ttccaccaaa ctctgcaaga
     1321 tcccagagtg agaggcctgt atttccctgc tggtggctcc agttcaggaa cagtaaaccc
     1381 tgttctgact actgcctctc ccttatcgtc aatcttctcg aggattgggg accctgcgct
     1441 gaacatggag aacatcacat caggattcct aggacccctt ctcgtgttac aggcggggtt
     1501 tttcttgttg acaagaatcc tcacaatacc gcagagtcta gactcgtggt ggacttctct
     1561 caattttcta gggggaacta ccgtgtgtct tggccaaaat tcgcagtccc caacctccaa
     1621 tcactcacca acctcttgtc ctccaacttg tcctggttat cgctggatgt gtctgcggcg
     1681 ttttatcatc ttcctcttca tcctgctgct atgcctcatc ttcttgttgg ttcttctgga
     1741 ctatcaaggt atgttgcccg tttgtcctct aattccagga tcctcaacaa ccagcacggg
     1801 accatgccgg acctgcatga ctactgctca aggaacctct atgtatccct cctgttgctg
     1861 taccaaacct tcggacggaa attgcacctg tattcccatc ccatcatcct gggctttcgg
     1921 aaaattccta tgggagtggg cctcagcccg tttctcctgg ctcagtttac tagtgccatt
     1981 tgttcagtgg ttcgtagggc tttcccccac tgtttggctt tcagttatat ggatgatgtg
     2041 gtattggggg ccaagtctgt acagcatctt gagtcccttt ttaccgctgt taccaatttt
     2101 cttttgtctt tgggtataca tttaagcggc cgctcgagca tgcatctaga gggccctatt
     2161 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     2221 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2281 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2341 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2401 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2461 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2521 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2581 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2641 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2701 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2761 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2821 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2881 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2941 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3001 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3061 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3181 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3241 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3301 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3361 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3421 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3481 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3541 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3601 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3661 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3721 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3781 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3841 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3901 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3961 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4021 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4081 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4141 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4201 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4261 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4321 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4381 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4441 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4501 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4561 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5041 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5281 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6601 tgacgtc
//
