LOCUS       Exported                6118 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses HBV Small envelope glycoprotein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    S-HBsAg
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6118)
  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 6118)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6118
                     /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             956..1636
                     /codon_start=1
                     /product="major/small (S) envelope protein of hepatitis B 
                     virus"
                     /label=HBsAg (S)
                     /note="hepatitis B surface antigen"
                     /translation="MENITSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGT
                     TVCLGQNSQSPTSNHSPTSCPPTCPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGML
                     PVCPLIPGSSTTSTGPCRTCMTTAQGTSMYPSCCCTKPSDGNCTCIPIPSSWAFGKFLW
                     EWASARFSWLSLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYSILSPFLPLLPIFFCL
                     WVYI"
     promoter        complement(1671..1689)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1672..1689)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1709..1726)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1715..1939
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1985..2413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2422..2442)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2427..2756
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2607..2742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2669..2688
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2823..3617
                     /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(2877..2896)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3487..3506
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3791..3912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3828..3847)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3882..3901
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3961..3977)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3961..3977)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3974..3996)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3985..4001
                     /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(4009..4039)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4054..4075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4192..4209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4363..4951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4443..4462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5122..5982)
                     /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     5745..5764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5983..6087)
                     /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 atcccatgga
      961 gaacatcaca tcaggattcc taggacccct tctcgtgtta caggcggggt ttttcttgtt
     1021 gacaagaatc ctcacaatac cgcagagtct agactcgtgg tggacttctc tcaattttct
     1081 agggggaact accgtgtgtc ttggccaaaa ttcgcagtcc ccaacctcca atcactcacc
     1141 aacctcttgt cctccaactt gtcctggtta tcgctggatg tgtctgcggc gttttatcat
     1201 cttcctcttc atcctgctgc tatgcctcat cttcttgttg gttcttctgg actatcaagg
     1261 tatgttgccc gtttgtcctc taattccagg atcctcaaca accagcacgg gaccatgccg
     1321 gacctgcatg actactgctc aaggaacctc tatgtatccc tcctgttgct gtaccaaacc
     1381 ttcggacgga aattgcacct gtattcccat cccatcatcc tgggctttcg gaaaattcct
     1441 atgggagtgg gcctcagccc gtttctcctg gctcagttta ctagtgccat ttgttcagtg
     1501 gttcgtaggg ctttccccca ctgtttggct ttcagttata tggatgatgt ggtattgggg
     1561 gccaagtctg tacagcatct tgagtccctt tttaccgctg ttaccaattt tcttttgtct
     1621 ttgggtatac atttaagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     1681 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1741 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1801 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1861 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     1921 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     1981 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2581 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2641 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2701 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2761 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     2821 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2881 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2941 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3001 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3061 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3121 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3181 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3241 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3301 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3361 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3421 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3481 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3541 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3601 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3661 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3721 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3781 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3841 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3901 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     3961 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4201 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4261 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4321 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4801 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6001 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6061 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
