LOCUS       Exported               14616 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCHIK-LR 5`GFP, complete sequence.
ACCESSION   EU224269
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14616)
  AUTHORS   Tsetsarkin K, Higgs S, McGee CE, De Lamballerie X, Charrel RN, 
            Vanlandingham DL.
  TITLE     Infectious clones of Chikungunya virus (La Reunion isolate) for 
            vector competence studies
  JOURNAL   Vector Borne Zoonotic Dis. 6 (4), 325-337 (2006)
  PUBMED    17187566
REFERENCE   2  (bases 1 to 14616)
  AUTHORS   Tsetsarkin KA, Vanlandingham DL, Higgs S.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-OCT-2007) Pathology, University of Texas Medical 
            Branch, University Blvd, Galveston, TX 77555-0609, USA
REFERENCE   3  (bases 1 to 14616)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14616
                     /organism="Cloning vector pCHIK-LR 5`GFP"
                     /mol_type="other DNA"
                     /note="derived from Chikungunya virus strain LR2006 OPY1 
                     and plasmid pSinRep5 which serves to propagate the viral 
                     cDNA in Escherichia coli"
                     /db_xref="taxon:481784"
     5UTR           1..76
     CDS             77..7501
                     /codon_start=1
                     /product="nonstructural polyprotein"
                     /label=nonstructural polyprotein
                     /note="encodes nonstructural proteins 1 through 4"
                     /protein_id="ABX40009.1"
                     /translation="MDPVYVDIDADSAFLKALQRAYPMFEVEPRQVTPNDHANARAFSH
                     LAIKLIEQEIDPDSTILDIGSAPARRMMSDRKYHCVCPMRSAEDPERLANYARKLASAA
                     GKVLDRNISGKIGDLQAVMAVPDTETPTFCLHTDVSCRQRADVAIYQDVYAVHAPTSLY
                     HQAIKGVRVAYWVGFDTTPFMYNAMAGAYPSYSTNWADEQVLKAKNIGLCSTDLTEGRR
                     GKLSIMRGKKLKPCDRVLFSVGSTLYPESRKLLKSWHLPSVFHLKGKLSFTCRCDTVVS
                     CEGYVVKRITMSPGLYGKTTGYAVTHHADGFLMCKTTDTVDGERMSFSVCTYVPATICD
                     QMTGILATEVTPEDAQKLLVGLNQRIVVNGRTQRNTNTMKNYLLPVVAQAFSKWAKECR
                     KDMEDEKLLGVRERTLTCCCLWAFKKQKTHTVYKRPDTQSIQKVQAEFDSFVVPSLWSS
                     GLSIPLRTRIKWLLSKVPKTDLIPYSGDAREARDAEKEAEEEREAELTREALPPLQAAQ
                     EDVQVEIDVEQLEDRAGAGIIETPRGAIKVTAQPTDHVVGEYLVLSPQTVLRSQKLSLI
                     HALAEQVKTCTHNGRAGRYAVEAYDGRVLVPSGYAISPEDFQSLSESATMVYNEREFVN
                     RKLHHIAMHGPALNTDEESYELVRAERTEHEYVYDVDQRRCCKKEEAAGLVLVGDLTNP
                     PYHEFAYEGLKIRPACPYKIAVIGVFGVPGSGKSAIIKNLVTRQDLVTSGKKENCQEIT
                     TDVMRQRGLEISARTVDSLLLNGCNRPVDVLYVDEAFACHSGTLLALIALVRPRQKVVL
                     CGDPKQCGFFNMMQMKVNYNHNICTQVYHKSISRRCTLPVTAIVSSLHYEGKMRTTNEY
                     NKPIVVDTTGSTKPDPGDLVLTCFRGWVKQLQIDYRGYEVMTAAASQGLTRKGVYAVRQ
                     KVNENPLYASTSEHVNVLLTRTEGKLVWKTLSGDPWIKTLQNPPKGNFKATIKEWEVEH
                     ASIMAGICSHQMTFDTFQNKANVCWAKSLVPILETAGIKLNDRQWSQIIQAFKEDKAYS
                     PEVALNEICTRMYGVDLDSGLFSKPLVSVYYADNHWDNRPGGKMFGFNPEAASILERKY
                     PFTKGKWNINKQICVTTRRIEDFNPTTNIIPANRRLPHSLVAEHRPVKGERMEWLVNKI
                     NGHHVLLVSGYNLALPTKRVTWVAPLGVRGADYTYNLELGLPATLGRYDLVVINIHTPF
                     RIHHYQQCVDHAMKLQMLGGDSLRLLKPGGSLLIRAYGYADRTSERVICVLGRKFRSSR
                     ALKPPCVTSNTEMFFLFSNFDNGRRNFTTHVMNNQLNAAFVGQVTRAGCAPSYRVKRMD
                     IAKNDEECVVNAANPRGLPGGGVCKAVYKKWPESFKNSATPVGTAKTVMCGTYPVIHAV
                     GPNFSNYSESEGDRELAAAYREVAKEVTRLGVNSVAIPLLSTGVYSGGKDRLTQSLNHL
                     FTAMDSTDADVVIYCRDKEWEKKISEAIQMRTQVELLDEHISIDCDIVRVHPDSSLAGR
                     KGYSTTEGALYSYLEGTRFHQTAVDMAEIHTMWPKQTEANEQVCLYALGESIESIRQKC
                     PVDDADASSPPKTVPCLCRYAMTPERVTRLRMNHVTSIIVCSSFPLPKYKIEGVQKVKC
                     SKVMLFDHNVPSRVSPREYRSSQESAQEASTITSLTHSQFDLSVDGEILPVPSDLDADA
                     PALEPALDDGATHTLPSTTGNLAAVSDWVMSTVPVAPPRRRRGRNLTVTCDEREGNITP
                     MASVRFFRAELCPVVQETAETRDTAMSLQAPPSTATEPNHPPISFGASSETFPITFGDF
                     NEGEIESLSSELLTFGDFLPGEVDDLTDSDWSTCSDTDDELRLDRAGGYIFSSDTGPGH
                     LQQKSVRQSVLPVNTLEEVHEEKCYPPKLDEAKEQLLLKKLQESASMANRSRYQSRKVE
                     NMKAAIIQRLKRGCRLYLMSETPKVPTYRTTYPAPVYSPPINVRLSNPESAVAACNEFL
                     ARNYPTVSSYQITDEYDAYLDMVDGSESCLDRATFNPSKLRSYPKQHAYHAPSIRSAVP
                     SPFQNTLQNVLAAATKRNCNVTQMRELPTLDSAVFNVECFKKFACNQEYWEEFAASPIR
                     ITTENLATYVTKLKGPKAAALFAKTHNLLPLQEVPMDRFTVDMKRDVKVTPGTKHTEER
                     PKVQVIQAAEPLATAYLCGIHRELVRRLNAVLLPNVHTLFDMSAEDFDAIIAAHFKPGD
                     TVLETDIASFDKSQDDSLALTALMLLEDLGVDHSLLDLIEAAFGEISSCHLPTGTRFKF
                     GAMMKSGMFLTLFVNTLLNITIASRVLEDRLTKSACAAFIGDDNIIHGVVSDELMAARC
                     ATWMNMEVKIIDAVVSLKAPYFCGGFILHDTVTGTACRVADPLKRLFKLGKPLAAGDEQ
                     DEDRRRALADEVIRWQRTGLIDELEKAVYSRYEVQGISVVVMSMATFASSRSNFEKLRG
                     PVITLYGGPK"
     regulatory      7480..7499
                     /regulatory_class="promoter"
                     /note="first subgenomic promoter"
     CDS             7569..8288
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=green fluorescent protein
                     /note="GFP"
                     /protein_id="ABX40008.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      8356..8375
                     /regulatory_class="promoter"
                     /note="second subgenomic promoter"
     CDS             8443..12189
                     /codon_start=1
                     /product="structural polyprotein"
                     /label=structural polyprotein
                     /protein_id="ABX40007.1"
                     /translation="MEFIPTQTFYNRRYQPRPWTPRPTIQVIRPRPRPQRQAGQLAQLI
                     SAVNKLTMRAVPQQKPRRNRKNKKQKQKQQAPQNNTNQKKQPPKKKPAQKKKKPGRRER
                     MCMKIENDCIFEVKHEGKVTGYACLVGDKVMKPAHVKGTIDNADLAKLAFKRSSKYDLE
                     CAQIPVHMKSDASKFTHEKPEGYYNWHHGAVQYSGGRFTIPTGAGKPGDSGRPIFDNKG
                     RVVAIVLGGANEGARTALSVVTWNKDIVTKITPEGAEEWSLAIPVMCLLANTTFPCSQP
                     PCTPCCYEKEPEETLRMLEDNVMRPGYYQLLQASLTCSPHRQRRSTKDNFNVYKATRPY
                     LAHCPDCGEGHSCHSPVALERIRNEATDGTLKIQVSLQIGIKTDDSHDWTKLRYMDNHM
                     PADAERAGLFVRTSAPCTITGTMGHFILARCPKGETLTVGFTDSRKISHSCTHPFHHDP
                     PVIGREKFHSRPQHGKELPCSTYVQSTAATTEEIEVHMPPDTPDRTLMSQQSGNVKITV
                     NGQTVRYKCNCGGSNEGLTTTDKVINNCKVDQCHAAVTNHKKWQYNSPLVPRNAELGDR
                     KGKIHIPFPLANVTCRVPKARNPTVTYGKNQVIMLLYPDHPTLLSYRNMGEEPNYQEEW
                     VMHKKEVVLTVPTEGLEVTWGNNEPYKYWPQLSTNGTAHGHPHEIILYYYELYPTMTVV
                     VVSVATFILLSMVGMAAGMCMCARRRCITPYELTPGATVPFLLSLICCIRTAKAATYQE
                     AAIYLWNEQQPLFWLQALIPLAALIVLCNCLRLLPCCCKTLAFLAVMSVGAHTVSAYEH
                     VTVIPNTVGVPYKTLVNRPGYSPMVLEMELLSVTLEPTLSLDYITCEYKTVIPSPYVKC
                     CGTAECKDKNLPDYSCKVFTGVYPFMWGGAYCFCDAENTQLSEAHVEKSESCKTEFASA
                     YRAHTASASAKLRVLYQGNNITVTAYANGDHAVTVKDAKFIVGPMSSAWTPFDNKIVVY
                     KGDVYNMDYPPFGAGRPGQFGDIQSRTPESKDVYANTQLVLQRPAVGTVHVPYSQAPSG
                     FKYWLKERGASLQHTAPFGCQIATNPVRAVNCAVGNMPISIDIPEAAFTRVVDAPSLTD
                     MSCEVPACTHSSDFGGVAIIKYAASKKGKCAVHSMTNAVTIREAEIEVEGNSQLQISFS
                     TALASAEFRVQVCSTQVHCAAECHPPKDHIVNYPASHTTLGVQDISATAMSWVQKITGG
                     VGLVVAVAALILIVVLCVSFSRH"
     3UTR           12190..12687
     promoter        12787..12891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             12892..13752
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      13923..14511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      14597..14616
                     /regulatory_class="promoter"
                     /note="Sp6 promoter"
     promoter        14599..1
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"