LOCUS       sequence-348681-        7236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HCV E1E2 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  AUTHORS   Salas JH, Urbanowicz RA, Guest JD, Frumento N, Figueroa A, Clark KE,
            Keck Z, Cowton VM, Cole SJ, Patel AH, Fuerst TR, Drummer HE, Major 
            M, Tarr AW, Ball JK, Law M, Pierce BG, Foung SKH, Bailey JR
  TITLE     An Antigenically Diverse, Representative Panel of Envelope 
            Glycoproteins for Hepatitis C Virus Vaccine Development.
  JOURNAL   Gastroenterology. 2022 Feb;162(2):562-574. doi: 
            10.1053/j.gastro.2021.10.005. Epub 2021 Oct 13.
  PUBMED    34655573
REFERENCE   2  (bases 1 to 7236)
  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: 
            "10.1053/j.gastro.2021.10.005"; journalName: "Gastroenterology"; 
            date: "2022-02"; volume: "162"; issue: "2"; pages: "562-574"
COMMENT     Sequence Label: 1b25
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    677..701
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2442..2466)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2492..2533
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2573..2592)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2617..2665
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2867..3295
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2954..2973)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3164..3185
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3304..3324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3309..3638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3489..3624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3551..3570
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3705..4499
                     /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(3759..3778)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4369..4388
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4675..4796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4712..4731)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4766..4785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4845..4861)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4845..4861)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4858..4880)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4869..4885
                     /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(4893..4899,4900..4917,4918..4923))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4938..4959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5076..5093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5247..5835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5327..5346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6006..6797,6798..6866))
                     /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     6629..6648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6867..6971)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7046..7065)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag ctatcaacaa gtttgtacaa aaaagcaggc ttcaccatgg gttgctcttt
      721 ctctatcttc ctcttggctt tgctgtcctg tttgaccatc ccagcttccg ctatcgaagt
      781 gcgcaacgtg tccggagtgt accatgtcac gaacgactgc tccaacgcaa gtattgtgta
      841 tgaggcagcg gacgtgatca tgcatacccc cgggtgcgtg ccctgcgtcc gggagggcaa
      901 ctcctcccgc tgctgggtag cgctcactcc cacgctcgcg gccaggaact ccagcatccc
      961 cactacgaca atacgacgcc acgtcgattt gctcgttggg acggctgctt tttgctccgc
     1021 tatgtacgtg ggagatcttt gcggatctgt tttccttgtc tctcagctgt tcaccttctc
     1081 gcctcgccgg catgagacag tacaggactg caattgttca atctatcccg gccacgtatc
     1141 tggtcaccgc atggcttggg acatgatgat gaactggtca cctacaacag ctctagtggt
     1201 gtcgcagtta ctccggatcc cacaagccgt cgtggacgtg gtggtgggag cccactgggg
     1261 agtcctggcg ggcctcgcct actactccat ggtggggaac tgggctaagg ttttgattgt
     1321 gatgctactt tttgccggtg tcgatggaga gacttacacg acgggggggg cgcaggcccg
     1381 taacgtccgc acgtttacga gcctcttcac ggctgggcca tcccaaaaaa tccagcttat
     1441 aaacaccaac ggcagttggc acatcaacag gactgccctg aactgcaatg actccctcaa
     1501 caccgggttc cttgccgcgc tgttctacgt ccagagattc aacgcgtccg gatgcccaga
     1561 gcgcatggcc agctgccgcc ccattgacaa attcgctcag gggtggggtc ccatcactta
     1621 tgttaagccg gccatctcgg accagaagcc ctattgttgg cactacgcac cccaaccgtg
     1681 cggtattgta cccgcgtcgc aggtgtgcgg tccagtgtac tgtttcaccc caagccctgt
     1741 tgtggtgggg acgaccgatc gtttcggcgt ccctacatat aactgggggg agaatgagac
     1801 ggacgtgctg cttctcaaca acacgcggcc gccacagggc aactggttcg gctgcacatg
     1861 gatgaatggc actgggttca ccaagacgtg cgggggcccc ccgtgcaaca tcgggggggc
     1921 cggtaatgac accttgacct gccccacgga ttgcttccgg aagcaccccg aggccactta
     1981 caccaaatgc ggttcggggc cctggctgac gcctagatgc atggttgact acccatacag
     2041 gctttggcat tacccctgca ctgtcaattt taccatcttc aaggttagga tgtatgtggg
     2101 gggtgtggaa cacaggctca ccgccgcatg caactggacc cgaggagagc gttgtgactt
     2161 ggaggacagg gatagatcag agcttagccc gctgctgctg tctacaacag agtggcaggt
     2221 gttgccctgt tccttcacca ccctaccggc tttgtccacc ggcttgatcc atctccaccg
     2281 gaacatcgtg gacgtgcaat acctgtacgg tatagggtca gcggttgtct cctacgcaat
     2341 taagtgggag tatgtcctgt tgctcttcct tttcctggcg gacgcgcgcg tctgtgcctg
     2401 cttgtggatg atgctgctaa tagctcaggc tgaggcctag gacccagctt tcttgtacaa
     2461 agtggttgat ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg
     2521 tctcgattct acgcgtaccg gttagtaatg agtttaaacg ggggaggcta actgaaacac
     2581 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     2641 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     2701 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     2761 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     2821 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     2881 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2941 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3001 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3061 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3121 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3181 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3241 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3301 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3361 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3481 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3541 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3601 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3661 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3721 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3781 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3841 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3901 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3961 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4021 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4081 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4141 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4201 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4261 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4321 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4381 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4441 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4501 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4561 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4621 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4681 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4741 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4801 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4861 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4921 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4981 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5041 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5101 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5161 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5221 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5281 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5341 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5401 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5461 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5521 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5581 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5641 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5701 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5761 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5821 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5881 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5941 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6001 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6061 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6121 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6181 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6241 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6301 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6361 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6421 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6481 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6541 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6601 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6661 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6721 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6781 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6841 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6901 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6961 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7021 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7081 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     7141 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     7201 cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
