LOCUS       Exported               12414 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pEZY202, complete sequence.
ACCESSION   EU181459
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12414)
  AUTHORS   Guo F, Wang Y, Zhang YZ.
  TITLE     Construction of two recombination yeast two-hybrid vectors by in 
            vitro recombination
  JOURNAL   Mol. Biotechnol. 36 (1), 38-43 (2007)
  PUBMED    17827536
REFERENCE   2  (bases 1 to 12414)
  AUTHORS   Guo F, Wang Y, Zhang Y-Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2007) Biological, Chemical, and Physical Sciences,
            Illinois Institute of Technology, 3101 South Dearborn St., Chicago, 
            IL 60616, USA
REFERENCE   3  (bases 1 to 12414)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12414
                     /organism="Cloning vector pEZY202"
                     /mol_type="other DNA"
                     /label=yeast two-hybrid vector
                     /note="yeast two-hybrid vector"
                     /db_xref="taxon:474433"
     regulatory      67..1511
                     /regulatory_class="promoter"
                     /note="ADH1 promoter"
     promoter        1110..1511
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_feature    1538..2144
                     /note="encodes partial LexA; contains DNA-binding domain"
     CDS             1538..2143
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5-CTGTATATATATACAG-3."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     misc_recomb     2156..2287
                     /label=attR1 site
                     /note="attR1 site"
     protein_bind    2156..2280
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2709..3014)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB lethal protein"
                     /function="gyrase inhibitor"
                     /label=ccdB
                     /protein_id="ABW37349.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(2709..3014)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(3359..4018)
                     /codon_start=1
                     /gene="CmR"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /protein_id="ABW37350.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(3359..4018)
                     /gene="CmR"
                     /label=CmR
     promoter        complement(4019..4121)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(4299..4423)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     4300..4423
                     /label=attR2 site
                     /note="attR2 site"
     terminator      4457..4644
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      4491..4786
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     rep_origin      5131..7061
                     /label=2 mu ori
                     /note="2 mu ori"
     rep_origin      5376..6718
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5708..5755
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(8362..9021)
                     /codon_start=1
                     /gene="HIS3"
                     /product="imidazoleglycerolphosphate dehydratase"
                     /label=HIS3
                     /protein_id="ABW37351.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     gene            complement(8362..9021)
                     /gene="HIS3"
                     /label=HIS3
     promoter        complement(9022..9209)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        10274..10378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             10379..11239
                     /codon_start=1
                     /gene="AmpR"
                     /product="ampicillin resistance"
                     /label=AmpR
                     /protein_id="ABW37352.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            10379..11239
                     /gene="AmpR"
                     /label=AmpR
     rep_origin      11381..12071
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      11410..11998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    12184..12324
                     /label=bom
                     /note="basis of mobility region from pBR322"