LOCUS       Exported                8960 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNG168, complete sequence.
ACCESSION   AY291460
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8960)
  AUTHORS   DasSarma S.
  TITLE     Natural plasmids and plasmid vectors of halophiles
  JOURNAL   (in) DasSarma,S., Fleischmann,E.M., Robb,F.T., Place,A.R., 
            Sowers,K.R. and Schreier,H.J. (Eds.); ARCHAEA, A LABORATORY MANUAL -
            HALOPHILES: 241-250; Cold Spring Harbor Laboratory Press, Cold 
            Spring Harbor, NY (1995)
REFERENCE   2  (bases 1 to 8960)
  AUTHORS   DasSarma S.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2003) Center of Marine Biotechnology, University 
            of Maryland Biotechnology Institute, 701 E. Pratt Street, Suite 236,
            Baltimore, MD 21202
REFERENCE   3  (bases 1 to 8960)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8960
                     /organism="Shuttle vector pNG168"
                     /lab_host="Halobacterium sp. NRC-1"
                     /mol_type="genomic DNA"
                     /note="Halobacterium sp. NRC-1 - Escherichia coli shuttle 
                     plasmid"
                     /db_xref="taxon:231449"
     CDS             849..3878
                     /codon_start=1
                     /gene="repH"
                     /product="RepH"
                     /label=repH
                     /note="replication protein; necessary for replication in 
                     haloarchaea"
                     /protein_id="AAP44985.1"
                     /translation="MHTPNQQQGIRKIVPGGTLSTAGITITEVTPRVTEWIPDLLEELL
                     PRSIQSVRKFIRQEDPEVLTHARYNTVYRRLQEETLRFDHQEWCSTTDIWSDAEAEAVE
                     YVESLVEFAVKYSDVDEDDLDELSEYHQQRCKSLKQTLTTISTGRGPLNAGLEALAKGP
                     VRLHDELDDAPQPITLVLDGELWSKLDDRGTGIRALAAIAVLGSTFDVRLVISPALDAA
                     IERRYPDWYDSHLRLTETRETSSVESAGGDGQPSAEQLEEAWEAIQNLPEESGRLRLLR
                     NLPIEGSRDYRDLKQDDEIDVQAGTVGRYILDLEELGLVDIDRRGQYNSASLTGLGQVA
                     VEQYVTTDYRVIHPTQSTLETHLTPTPQPQASTVYPARSDTREGDQPGTAEDWIAATGS
                     PSEGADYVQWLDGPSGVLDAWGMHQRYLAGRRDRGVTLVDDRIERFEDGRVSYLSCFDD
                     DLFVATQWGGPLPTLGRIAGALLSDKALSKILTPSRLGNQFEEINDAVVEQLDREAGEI
                     IRRGHQIGWFSEDEEDYDGWRERIGSVRSLCLQQVGELTNSDDVEARTELLRDLHGLVA
                     SATQLYYAAGVDVTINVRVPDTGMLISDERRLDDFLGFARYTIPKQSVYGIHSGYRMLL
                     EDRPEKLKRRLPYEVDDADSTMHLTASWVFSGSTMIDLHDDIEDAIEMETNEIREAIAN
                     GQESAPVMEIPVQIGNSYSAIRNHVEDYASAKNYQVAHQEDIHEGKQDLERLVRLFLRV
                     LGTEDRPHRACPHDVAEAMLHVAQSSRNYDFITVRDISYGLSNLPTKRLLPELPPTATK
                     LLKTLLDADDPMGRSEIIDTADISESSYDRYINELAAWDIIEPREIEGHRRWEAHLEPW
                     WTPQSDRDEPYADPDPDTGILYAEFPRDVASAVMCHLITHYDLPDLETAYLEGIQPGDD
                     IKALFDDHDRLRRWRPFLWGAFADSDKLERGPSGTAASDSTVVRLGQSPGPDTAQSSFQ
                     DVSETATQRDRLSQPSPGLD"
     gene            849..3878
                     /gene="repH"
                     /label=repH
     protein_bind    4431..4452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4467..4497
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4505..4521
                     /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)."
     primer_bind     4529..4545
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4541..5116
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="LacZalpha"
                     /label=lacZalpha
                     /note="LacZ alpha fragment; blue-white screening"
                     /protein_id="AAP44986.1"
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     gene            4541..5116
                     /gene="lacZalpha"
                     /label=lacZalpha
     promoter        4566..4584
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    4597..4704
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     misc_feature    4603..4674
                     /note="multiple cloning region: ApaI, HindIII, EcoRI, PstI,
                     SmaI, BamHI, SpeI, NotI"
     primer_bind     4614..4630
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4664..4680)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4713..4731)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4741..4757)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4899..5354
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5380..5484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5485..6345
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="confers resistance to ampicillin"
                     /label=bla
                     /note="Bla"
                     /protein_id="AAP44987.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5485..6345
                     /gene="bla"
                     /label=bla
     rep_origin      6516..7104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7425..8636)
                     /codon_start=1
                     /product="3-hydroxy-3-methylglutaryl-coenzyme A reductase"
                     /function="confers resistance to mevinolin"
                     /label=3-hydroxy-3-methylglutaryl-coenzyme A reductase
                     /note="Hmg"
                     /protein_id="AAP44988.1"
                     /translation="MTDAASLADRVREGDLRLHELEAHADADTAAEARRLLVESQSGAS
                     LDAVGNYGFPAEAAESAIENMVGSIQVPMGVAGPVSVDGGSVAGEKYLPLATTEGALLA
                     SVNRGCSVINSAGGATARVLKSGMTRAPVFRVADVAEAEALVSWTRDNFAALKEAAEET
                     TNHGELLDVTPYVVGNSVYLRFRYDTKDAMGMNMATIATEAVCGVVEAETAASLVALSG
                     NLCSDKKPAAINAVEGRGRSVTADVRIPREVVEERLHTTPERGRELNTRKNLVGSAKAA
                     SLGFNAHVANVVAAMFLATGQDEAQVVEGANAITTAEVQDGDLYVSVSIASLEVGTVGG
                     GTKLPTQSEGLDILGVSGGGDPAGSNADALAECIAVGSLAGELSLLSALASRHLSSAHA
                     ELGR"