LOCUS       sequence-189896-        7259 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of HCV E1E2 genes cloned from patient serum. United 
            Kingdom Nottingham Panel x.xx.xx (genotype.patient number.clone 
            number).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  AUTHORS   Urbanowicz RA, McClure CP, Brown RJ, Tsoleridis T, Persson MA, Krey 
            T, Irving WL, Ball JK, Tarr AW
  TITLE     A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in 
            Vaccine Research Reveals Extremes of Monoclonal Antibody 
            Neutralization Resistance.
  JOURNAL   J Virol. 2015 Dec 23;90(7):3288-301. doi: 10.1128/JVI.02700-15.
  PUBMED    26699643
REFERENCE   2  (bases 1 to 7259)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1128/JVI"; 
            journalName: "J Virol"; date: "2015-12-23- 23"; volume: "90"; issue:
            "7"; pages: "3288-301"
COMMENT     Sequence Label: UKNP1.20.3
FEATURES             Location/Qualifiers
     source          1..7259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..57)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        229..608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        609..812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     763..783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     857..876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        857..875
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2751..2792
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2802..2819
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2842..2859)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2848..3072
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3118..3546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3205..3224)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3415..3436
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3555..3575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3560..3889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3740..3875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3802..3821
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3956..4750
                     /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(4010..4029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4620..4639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4926..5047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4963..4982)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5017..5036
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5096..5112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5096..5112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5109..5131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5120..5136
                     /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(5144..5150,5151..5168,5169..5174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5189..5210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5327..5344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5498..6086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5578..5597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6257..7048,7049..7117))
                     /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     6880..6899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7118..7222)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
       61 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
      121 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
      181 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      241 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      301 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      361 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      421 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      481 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      541 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      601 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      661 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      721 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      781 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      841 actggcttat cgaaattaat acgactcact atagggagac ccaagctggc tagttaagct
      901 tggtaccgag ctcggatcca gtacccttca ccatgggttg ctctttctct atcttcctct
      961 tggccttgct gtcctgcctg accatcccag cttccgctta tgaggtgcgc aacgcgtccg
     1021 gggtgtacca tgtcacgaac gactgctcca actcaagtat tgtgtatgag gcagcggaca
     1081 tgatcatgca catccctggg tgcgtgcctt gcgttcggga gagcaacctc tcccgctgct
     1141 gggtagcgct cactcccacg ctcgcggcca ggaatgccac cgtccctact acggcgatac
     1201 gacgtcatgt cgatttgctt gttggggcgg ccgctttctg ctctgccatg tacgtggggg
     1261 atctctgcgg atctgttctc cttgtttccc agctgttcac cttctcgcct cgtcggcatg
     1321 agacggtgca ggactgcaat tgttcgatct atcccggtta cgtgtcaggt caccgcatgg
     1381 cttgggatat gatgatgaac tggtcaccta caacagctct agtggtgtcg cagttactcc
     1441 ggatcccaca agctgtcgtg gacatggtgg cgggagccca ctggggagtc ctagcgggcc
     1501 ttgcctacta ttccatggtg gggaactggg ctaaggtttt gattgtgatg ctgctctttg
     1561 ccggcgttga cgggaaaacc actgtggtgg gggggacgca aagccgcgcc ctccaaggcc
     1621 ttacgagcct tttcagcccg gggtcgcgtc agaatatcca gcttataaac accaatggca
     1681 gttggcacat caacaggact gctctgaact gtaatgactc cctcaacacc gggttcattg
     1741 ccgcgctgtt ctacacacac aaattcaact catccggatg cccagagcgc atggccagct
     1801 gccgctccat cgacaagttt gatcaagggt ggggtcccat cacctatgct gcgtcccgta
     1861 gtttggacca gaggccctat tgctggcatt atgcacctga accgtgtggt actgtacctg
     1921 cgacgcaggt gtgcggcccc gtgtattgtt tcactccaag ccctgttgtg gtggggacga
     1981 ccgatcgctt cggcgctcct acgtataact ggggggagaa tgagacagat gtgttgatcc
     2041 tcaacagcac gcggccgccg caaggtagtt ggtttgggtg cacatggatg aatggcactg
     2101 ggttcaccaa gacatgcggg ggccctccgt gcaacatcgg gggggtcggc aataatacct
     2161 tgacctgccc cacggactgc ttccggaagc accccgaggc cacttacacc agatgcggct
     2221 cggggccttg gttgacacct aggtgcatgg ttgactaccc atacaggctc tggcattacc
     2281 cctgcactgt caacttttcc atctttaagg tgagaatgta tgtggggggc gtggaacata
     2341 ggttcaacgc tgcatgcaat tggactcgag gcgaacgctg tgacttggag gacagggata
     2401 gatcagagct cagcccgctg ctgctatcca caacagagtg gcaggtgctg ccctgttcct
     2461 tcgccacctt accggcgctg tccacagggt tgatccatct ccatcagaac accgtggacg
     2521 tgcaatatct gtacggtata gggtcagcgg ttgtctccta tgcaatcaga tgggagtatg
     2581 tcctgttgct cttccttcta ctggcggacg cgcgcgtctg cgcctgcctg tggatgatgc
     2641 tgctgatatc ccaagctgag gcctaatcta gaaactttaa gggtcaagac aattctgcag
     2701 atatccagca cagtggcggc cgctcgagtc tagagggccc gcggttcgaa ggtaagccta
     2761 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt
     2821 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2881 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2941 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3001 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3061 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3121 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3181 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3241 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3301 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3361 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3421 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3481 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3541 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3661 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3721 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3781 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3841 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3901 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3961 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4021 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4081 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4141 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4201 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4261 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4321 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4381 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4441 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4501 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4561 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4621 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4681 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4741 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc
     4801 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4861 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4921 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4981 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5041 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5101 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5161 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5221 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5281 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5341 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5401 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5461 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5521 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5581 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5641 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5701 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5761 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5821 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5881 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5941 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6001 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6061 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6121 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6181 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6241 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6301 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6361 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6421 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6481 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6541 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6601 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6661 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6721 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6781 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6841 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6901 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6961 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7021 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7081 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7141 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7201 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacgga
//
