LOCUS       Exported                9655 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGR8, complete sequence.
ACCESSION   AD001531
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9655)
  AUTHORS   Mazzarella R, Pilia G.
  TITLE     Recombination trapping: an in vivo approach to recover cDNAs 
            encoded in YACs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9655)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     GSDB:S:1274444.
FEATURES             Location/Qualifiers
     source          1..9655
                     /organism="Cloning vector pGR8"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:64712"
     misc_feature    1..135
                     /standard_name="telomere"
                     /label=telomere
     prim_transcript 145..2097
                     /gene="HIS5"
     gene            145..2097
                     /gene="HIS5"
                     /label=HIS5
     CDS             761..1915
                     /codon_start=1
                     /gene="HIS5"
                     /product="histidinol phosphate aminotransferase"
                     /EC_number="2.6.1.9"
                     /label=HIS5
                     /protein_id="AAB68950.1"
                     /translation="MVFDLKRIVRPKIYNLEPYRCARDDFTEGILLDANENAHGPTPVE
                     LSKTNLHRYPDPHQLEFKTAMTKYRNKTSSYANDPEVKPLTADNLCLGVGSDESIDAII
                     RACCVREEKILVLPPTYSMSSVCANINDIEVVQCPLIVSDGSFQMDTEAVLTILKNDSL
                     IKLMFVTSPGNPTGAKIKTSLIEKVLQNWDNGLVVVDEAYVDFCGGSTAPLVTKYPNLV
                     TLQTLSKSFGLAGIRLGMTYATAELARILNAMKAPYNISSLASEYALKAVQDSNLKKME
                     ATSKIINEEKMLLLKELTALDYVDDQYVGGLDANFLLIRINGGDNVLAKKLYYQLATQS
                     GVVVRFRGNELGCSGCLRITVGTHEENTHLIKYFKETLYKLANE"
     promoter        complement(2100..2118)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     prim_transcript 2138..6708
                     /gene="LYS2"
     gene            2138..6708
                     /gene="LYS2"
                     /label=LYS2
     CDS             2374..6552
                     /codon_start=1
                     /gene="LYS2"
                     /product="alpha-aminoadipate reductase"
                     /EC_number="1.2.1.31"
                     /label=LYS2
                     /protein_id="AAB68951.1"
                     /translation="MTNEKVWIEKLDNPTLSVLPHDFLRPQQEPYTKQATYSLQLPQLD
                     VPHDSFSNKYAVALSVWAALIYRVTGDDDIVLYIANNKILRFNIQPTWSFNELYSTINN
                     ELNKLNSIEANFSFDELAEKIQSCQDLERTPQLFRLAFLENQDFKLDEFKHHLVDFALN
                     LDTSNNAHVLNLIYNSLLYSNERVTIVADQFTQYLTAALSDPSNCITKISLITASSKDS
                     LPDPTKNLGWCDFVGCIHDIFQDNAEAFPERTCVVETPTLNSDKSRSFTYRDINRTSNI
                     VAHYLIKTGIKRGDVVMIYSSRGVDLMVCVMGVLKAGATFSVIDPAYPPARQTIYLGVA
                     KPRGLIVIRAAGQLDQLVEDYINDELEIVSRINSIAIQENGTIEGGKLDNGEDVLAPYD
                     HYKDTRTGVVVGPDSNPTLSFTSGSEGIPKGVLGRHFSLAYYFNWMSKRFNLTENDKFT
                     MLSGIAHDPIQRDMFTPLFLGAQLYVPTQDDIGTPGRLAEWMSKYGCTVTHLTPAMGQL
                     LTAQATTPFPKLHHAFFVGDILTKRDCLRLQTLAENCRIVNMYGTTETQRAVSYFEVKS
                     KNDDPNFLKKLKDVMPAGKGMLNVQLLVVNRNDRTQICGIGEIGEIYVRAGGLAEGYRG
                     LPELNKEKFVNNWFVEKDHWNYLDKDNGEPWRQFWLGPRDRLYRTGDLGRYLPNGDCEC
                     CGRADDQVKIRGFRIELGEIDTHISQHPLVRENITLVRKNADNEPTLITFMVPRFDKPD
                     DLSKFQSDVPKEVETDPIVKGLIGYHLLSKDIRTFLKKRLASYAMPSLIVVMDKLPLNP
                     NGKVDKPKLQFPTPKQLNLVAENTVSETDDSQFTNVEREVRDLWLSILPTKPASVSPDD
                     SFFDLGGHSILATKMIFTLKKKLQVDLPLGTIFKYPTIKAFAAEIDRIKSSGGSSQGEV
                     VENVTANYAEDAKKLVETLPSSYPSREYFVEPNSAEGKTTINVFVTGVTGFLGSYILAD
                     LLGRSPKNYSFKVFAHVRAKDEEAAFARLQKAGITYGTWNEKFASNIKVVLGDLSKSQF
                     GLSDEKWMDLANTVDIIIHNGALVHWVYPYAKLRDPNVISTINVMSLAAVGKPKFFDFV
                     SSTSTLDTEYYFNLSDKLVSEGKPGILESDDLMNSASGLTGGYGQSKWAAEYIIRRAGE
                     RGLRGCIVRPGYVTGASANGSSNTDDFLLRFLKGSVQLGKIPDIENSVNMVPVDHVARV
                     VVATSLNPPKENELAVAQVTGHPRILFKDYLYTLHDYGYDVEIESYSKWKKSLEASVID
                     RNEENALYPLLHMVLDNLPESTKAPELDDRNAVASLKKDTAWTGVDWSNGIGVTPEEVG
                     IYIAFLNKVGFLPPPTHNDKLPLPSIELTQAQISLVASGAGARGSSAAA"
     misc_feature    complement(6709..7185)
                     /standard_name="f(+) origin"
                     /label=f(+) origin
     rep_origin      complement(6719..7147)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(6930..7502)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAGF
                     PRQALNRGLPLGFRFSALRHLDPKKLD"
     prim_transcript 7186..7496
                     /gene="LacZ"
     gene            7186..7496
                     /gene="LacZ"
                     /label=LacZ
     primer_bind     7289..7305
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7312..7330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7339..7446
                     /gene="LacZ"
                     /standard_name="multiple cloning sites"
                     /label=multiple cloning sites
     primer_bind     7363..7379
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7413..7429)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7459..7477)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7498..7514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7522..7538
                     /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(7546..7576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7591..7612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7900..8488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(7910..8030)
                     /direction=LEFT
                     /standard_name="plasmid ORI"
                     /label=plasmid ORI
     gene            complement(8659..9522)
                     /gene="AMP"
                     /label=AMP
     CDS             complement(8659..9519)
                     /codon_start=1
                     /gene="AMP"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=AMP
                     /protein_id="AAB68952.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     prim_transcript complement(8734..9522)
                     /gene="AMP"
     promoter        complement(9520..9624)
                     /gene="bla"
                     /label=AmpR promoter