pCHIKV-LS3 vector (V008519)

Basic Vector Information

Vector Name:
pCHIKV-LS3
Antibiotic Resistance:
Ampicillin
Length:
14532 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Scholte FE, Tas A, Martina BE, Cordioli P, Narayanan K, Makino S, Snijder EJ, van Hemert MJ.

pCHIKV-LS3 vector Map

pCHIKV-LS314532 bp7001400210028003500420049005600630070007700840091009800105001120011900126001330014000M13 fwd5'UTRnonstructural polyproteinStructural polyprotein3'UTRM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

pCHIKV-LS3 vector Sequence

LOCUS       V008519                14532 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008519
VERSION     V008519
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14532)
  AUTHORS   Scholte FE, Tas A, Martina BE, Cordioli P, Narayanan K, Makino S,
            Snijder EJ, van Hemert MJ.
  TITLE     Characterization of Synthetic Chikungunya Viruses Based on the
            Consensus Sequence of Recent E1-226V Isolates
  JOURNAL   PLoS ONE 8 (8), E71047 (2013)
   PUBMED   23936484
REFERENCE   2  (bases 1 to 14532)
  AUTHORS   Scholte FEM., Tas A, Snijder EJ, van Hemert MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-OCT-2012) Medical Microbiology, Leiden University
            Medical Center, Albinusdreef 2, Leiden, Zuid-Holland 2333 ZA, The
            Netherlands
REFERENCE   3  (bases 1 to 14532)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 14532)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
            date: "2013"; volume: "8"; issue: "8"; pages: "E71047"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (07-OCT-2012) Medical Microbiology, Leiden University Medical
            Center, Albinusdreef 2, Leiden, Zuid-Holland 2333 ZA, The
            Netherlands"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      410..430
                     /regulatory_class="promoter"
     5'UTR           430..505
     CDS             506..7930
                     /codon_start=1
                     /transl_except=(pos: 6074 .. 6076,  aa: OTHER)
                     /product="nonstructural polyprotein"
                     /label="nonstructural polyprotein"
                     /note="encodes nonstructural proteins 1 through 4"
                     /protein_id="AGN96196.1"
                     /translation="MDPVYVDIDADSAFLKALQRAYPMFEVEPRQVTPNDHANARAFSH
                     LAIKLIEQEIDPDSTILDIGSAPARRMMSDRKYHCVCPMRSAEDPERLANYARKLASAA
                     GKVLDRNISGKIGDLQAVMAVPDKETPTFCLHTDVSCRQRADVAIYQDVYAVHAPTSLY
                     HQAIKGVRVAYWVGFDTTPFMYNAMAGAYPSYSTNWADEQVLKAKNIGLCSTDLTEGRR
                     GKLSIMRGKKLKPCDRVLFSVGSTLYPESRKLLKSWHLPSVFHLKGKLSFTCRCDTVVS
                     CEGYVVKRITMSPGLYGKTTGYAVTHHADGFLMCKTTDTVDGERVSFSVCTYVPATICD
                     QMTGILATEVTPEDAQKLLVGLNQRIVVNGRTQRNMNTMKNYLLPVVAQAFSKWAKECR
                     KDMEDEKLLGVRERTLTCCCLWAFKKQKTHTVYKRPDTQSIQKVQAEFDSFVVPSLWSS
                     GLSIPLRTRIKWLLSKVPKTDLIPYSGDAREARDAEKEAEEEREAELTREALPPLQAAQ
                     EDVQVEIDVEQLEDRAGAGIIETPRGAIKVTAQPTDHVVGEYLVLSPQTVLRSQKLSLI
                     HALAEQVKTCTHNGRAGRYAVEAYDGRVLVPSGYAISPEDFQSLSESATMVYNEREFVN
                     RKLHHIAMHGPALNTDEESYELVRAERTEHEYVYDVDQRRCCKKEEAAGLVLVGDLTNP
                     PYHEFAYEGLKIRPACPYKIAVIGVFGVPGSGKSAIIKNLVTRQDLVTSGKKENCQEIT
                     TDVMRQRGLEISARTVDSLLLNGCNRPVDVLYVDEAFACHSGTLLALIALVRPRQKVVL
                     CGDPKQCGFFNMMQMKVNYNHNICTQVYHKSISRRCTLPVTAIVSSLHYEGKMRTTNEY
                     NKPIVVDTTGSTKPDPGDLVLTCFRGWVKQLQIDYRGYEVMTAAASQGLTRKGVYAVRQ
                     KVNENPLYASTSEHVNVLLTRTEGKLVWKTLSGDPWIKTLQNPPKGNFKATIKEWEVEH
                     ASIMAGICSHQMTFDTFQNKANVCWAKSLVPILETAGIKLNDRQWSQIIQAFKEDKAYS
                     PEVALNEICTRMYGVDLDSGLFSKPLVSVYYADNHWDNRPGGKMFGFNPEAASILERKY
                     PFTKGKWNINKQICVTTRRIEDFNPTTNIIPANRRLPHSLVAEHRPVKGERMEWLVNKI
                     NGHHVLLVSGYNLALPTKRVTWVAPLGVRGADYTYNLELGLPATLGRYDLVVINIHTPF
                     RIHHYQQCVDHAMKLQMLGGDSLRLLKPGGSLLIRAYGYADRTSERVICVLGRKFRSSR
                     ALKPPCVTSNTEMFFLFSNFDNGRRNFTTHVMNNQLNAAFVGQVTRAGCAPSYRVKRMD
                     IAKNDEECVVNAANPRGLPGDGVCKAVYKKWPESFKNSATPVGTAKTVMCGTYPVIHAV
                     GPNFSNYSESEGDRELAAAYREVAKEVTRLGVNSVAIPLLSTGVYSGGKDRLTQSLNHL
                     FTAMDSTDADVVIYCRDKEWEKKISEAIQMRTQVELLDEHISIDCDIVRVHPDSSLAGR
                     KGYSTTEGALYSYLEGTRFHQTAVDMAEIHTMWPKQTEANEQVCLYALGESIESIRQKC
                     PVDDADASSPPKTVPCLCRYAMTPERVTRLRMNHVTSIIVCSSFPLPKYKIEGVQKVKC
                     SKVMLFDHNVPSRVSPREYRSSQESAQEASTITSLTHSQFDLSVDGEILPVPSDLDADA
                     PALEPALDDGATHTLPSTTGNLAAVSDWVMSTVPVAPPRRRRGRNLTVTCDEREGNITP
                     MASVRFFRAELCPVVQETAETRDTAMSLQAPPSTATEPNHPPISFGASSETFPITFGDF
                     NEGEIESLSSELLTFGDFLPGEVDDLTDSDWSTCSDTDDEL*LDRAGGYIFSSDTGPGH
                     LQQKSVRQSVLPVNTLEEVHEEKCYPPKLDEAKEQLLLKKLQESASMANRSRYQSRKVE
                     NMKAAIIQRLKRGCRLYLMSETPKVPTYRTTYPAPVYSPPINVRLSNPESAVAACNEFL
                     ARNYPTVSSYQITDEYDAYLDMVDGSESCLDRATFNPSKLRSYPKQHAYHAPSIRSAVP
                     SPFQNTLQNVLAAATKRNCNVTQMRELPTLDSAVFNVECFKKFACNQEYWEEFAASPIR
                     ITTENLATYVTKLKGPKAAALFAKTHNLLPLQEVPMDRFTVDMKRDVKVTPGTKHTEER
                     PKVQVIQAAEPLATAYLCGIHRELVRRLNAVLLPNVHTLFDMSAEDFDAIIAAHFKPGD
                     TVLETDIASFDKSQDDSLALTALMLLEDLGVDHSLLDLIEAAFGEISSCHLPTGTRFKF
                     GAMMKSGMFLTLFVNTLLNITIASRVLEDRLTKSACAAFIGDDNIIHGVVSDELMAARC
                     ATWMNMEVKIIDAVVSLKAPYFCGGFILHDTVTGTACRVADPLKRLFKLGKPLAAGDEQ
                     DEDRRRALADEVIRWQRTGLIDELEKAVYSRYEVQGISVVVMSMATFASSRSNFEKLRG
                     PVITLYGGPK"
     regulatory      7909..7928
                     /regulatory_class="promoter"
     CDS             7996..11739
                     /note="Structural polyprotein from Chikungunya virus
                     (strain S27-African prototype). Accession#: Q8JUX5"
                     /label="Structural polyprotein"
     3'UTR           11743..12280
     primer_bind     complement(12311..12327)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    12335..12351
                     /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(12359..12389)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(12404..12425)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(12713..13301)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(13475..14332)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(14333..14437)
                     /label="AmpR promoter"

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