LOCUS       40924_19006        8718 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pEZY45, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8718)
  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 8718)
  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 8718)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8718)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. 
            Biotechnol."; date: "2007"; volume: "36"; issue: "1"; pages: "38-43"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-SEP-2007) Biological, Chemical, and Physical Sciences, Illinois 
            Institute of Technology, 3101 South Dearborn St., Chicago, IL 60616,
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        81..522
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     CDS             546..566
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             573..809
                     /codon_start=1
                     /label=B42 transcriptional activator
                     /note="yeast transcriptional activator cretaed from E. coli
                     genomic DNA fragments (Ma and Ptashne, 1987)"
                     /translation="INKDIEECNAIIEQFIDYLRTGQEMPMEMADQAINVVPGMTPKTI
                     LHAGPPIQPDWLKSNGFHEIEADVNDTSLLLSGD"
     CDS             816..842
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     protein_bind    861..985
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1417..1719)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2067..2723)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2724..2826)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3004..3128)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3273..3460
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      3982..5324
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5688..6359)
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(6360..6461)
                     /label=TRP1 promoter
     promoter        6564..6668
                     /label=AmpR promoter
     CDS             6669..7526
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7700..8288
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8576..8597
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8612..8642
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8650..8666
                     /label=lac operator
                     /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     8674..8690
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccccattatc ttagcctaaa aaaaccttct ctttggaact ttcagtaata cgcttaactg
       61 ctcattgcta tattgaagta cggattagaa gccgccgagc gggtgacagc cctccgaagg
      121 aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc
      181 tcgcgccgca ctgctccgaa caataaagat tctacaatac tagcttttat ggttatgaag
      241 aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa tgaacgaatc aaattaacaa
      301 ccataggatg ataatgcgat tagtttttta gccttatttc tggggtaatt aatcagcgaa
      361 gcgatgattt ttgatctatt aacagatata taaatgcaaa aactgcataa ccactttaac
      421 taatactttc aacattttcg gtttgtatta cttcttattc aaatgtaata aaagtatcaa
      481 caaaaaattg ttaatatacc tctatacttt aacgtcaagg aggaattaag cttatgggtg
      541 ctcctccaaa aaagaagaga aaggtagctg gtatcaataa agatatcgag gagtgcaatg
      601 ccatcattga gcagtttatc gactacctgc gcaccggaca ggagatgccg atggaaatgg
      661 cggatcaggc gattaacgtg gtgccgggca tgacgccgaa aaccattctt cacgccgggc
      721 cgccgatcca gcctgactgg ctgaaatcga atggttttca tgaaattgaa gcggatgtta
      781 acgataccag cctcttgctg agtggagatg cctcctaccc ttatgatgtg ccagattatg
      841 cctctcccga attcattaat acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg
      901 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
      961 aaacacaaca tatccagtca ctatgaatca actacttaga tggtattagt gacctgtagt
     1021 cgaccgacag ccttccaaat gttcttcggg tgatgctgcc aacttagtcg accgacagcc
     1081 ttccaaatgt tcttctcaaa cggaatcgtc gtatccagcc tactcgctat tgtcctcaat
     1141 gccgtattaa atcataaaaa gaaataagaa aaagaggtgc gagcctcttt tttgtgtgac
     1201 aaaataaaaa catctaccta ttcatatacg ctagtgtcat agtcctgaaa atcatctgca
     1261 tcaagaacaa tttcacaact cttatacttt tctcttacaa gtcgttcggc ttcatctgga
     1321 ttttcagcct ctatacttac taaacgtgat aaagtttctg taatttctac tgtatcgacc
     1381 tgcagactgg ctgtgtataa gggagcctga catttatatt ccccagaaca tcaggttaat
     1441 ggcgtttttg atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga
     1501 gaccggcaca ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac
     1561 caccgggtaa agttcacggg agactttatc tgacagcaga cgtgcactgg ccagggggat
     1621 caccatccgt cgcccgggcg tgtcaataat atcactctgt acatccacaa acagacgata
     1681 acggctctct cttttatagg tgtaaacctt aaactgcatt tcaccagccc ctgttctcgt
     1741 cagcaaaaga gccgttcatt tcaataaacc gggcgacctc agccatccct tcctgatttt
     1801 ccgctttcca gcgttcggca cgcagacgac gggcttcatt ctgcatggtt gtgcttacca
     1861 gaccggagat attgacatca tatatgcctt gagcaactga tagctgtcgc tgtcaactgt
     1921 cactgtaata cgctgcttca tagcatacct ctttttgaca tacttcgggt atacatatca
     1981 gtatatattc ttataccgca aaaatcagcg cgcaaatacg catactgtta tctggctttt
     2041 agtaagccgg atccacgcgg cgtttacgcc ccgccctgcc actcatcgca gtactgttgt
     2101 aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg aacctgaatc
     2161 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     2221 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     2281 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     2341 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     2401 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     2461 acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca
     2521 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     2581 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     2641 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     2701 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     2761 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     2821 cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca
     2881 ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt
     2941 gcgtcgggtg atgctgccaa cttagtcgac tacaggtcac taataccatc taagtagttg
     3001 attcatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa
     3061 atctaattta atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac
     3121 aaagtggtcc tcgagaagct ttggacttct tcgccagagg tttggtcaag tctccaatca
     3181 aggttgtcgg cttgtctacc ttgccagaaa tttacgaaaa gatggaaaag ggtcaaatcg
     3241 ttggtagata cgttgttgac acttctaaat aagcgaattt cttatgattt atgattttta
     3301 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     3361 taaaacgaaa attcttgttc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     3421 tatagcatga ggtcgctctt attgaccaca cctctaccgg catgccgagc aaatgcctgc
     3481 aaatcgctcc ccatttcacc caattgtaga tatgctaact ccagcaatga gttgatgaat
     3541 ctcggtgtgt attttatgtc ctcagaggac aacacctgtt gtaatcgttc ttccacacgg
     3601 atcctctaga gtcgactagc ggccgcttcg acctgcagca attctgaacc agtcctaaaa
     3661 cgagtaaata ggaccggcaa ttcttcaagc aataaacagg aataccaatt attaaaagat
     3721 aacttagtca gatcgtacaa taaagctttg aagaaaaatg cgccttattc aatctttgct
     3781 ataaaaaatg gcccaaaatc tcacattgga agacatttga tgacctcatt tctttcaatg
     3841 aagggcctaa cggagttgac taatgttgtg ggaaattgga gcgataagcg tgcttctgcc
     3901 gtggccagga caacgtatac tcatcagata acagcaatac ctgatcacta cttcgcacta
     3961 gtttctcggt actatgcata tgatccaata tcaaaggaaa tgatagcatt gaaggatgag
     4021 actaatccaa ttgaggagtg gcagcatata gaacagctaa agggtagtgc tgaaggaagc
     4081 atacgatacc ccgcatggaa tgggataata tcacaggagg tactagacta cctttcatcc
     4141 tacataaata gacgcatata agtacgcatt taagcataaa cacgcactat gccgttcttc
     4201 tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt
     4261 atgtgcgcag ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc tttgaagttc
     4321 ctattccgaa gttcctattc tctagaaagt ataggaactt cagagcgctt ttgaaaacca
     4381 aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt aacgagctac
     4441 taaaatattg cgaataccgc ttccacaaac attgctcaaa agtatctctt tgctatatat
     4501 ctctgtgcta tatccctata taacctaccc atccaccttt cgctccttga acttgcatct
     4561 aaactcgacc tctacatttt ttatgtttat ctctagtatt actctttaga caaaaaaatt
     4621 gtagtaagaa ctattcatag agtgaatcga aaacaatacg aaaatgtaaa catttcctat
     4681 acgtagtata tagagacaaa atagaagaaa ccgttcataa ttttctgacc aatgaagaat
     4741 catcaacgct atcactttct gttcacaaag tatgcgcaat ccacatcggt atagaatata
     4801 atcggggatg cctttatctt gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg
     4861 cgggaagtgg agtcaggctt tttttatgga agagaaaata gacaccaaag tagccttctt
     4921 ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag actgcattat agagcgcaca
     4981 aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa tagcgctctc gggatgcatt
     5041 tttgtagaac aaaaaagaag tatagattct ttgttggtaa aatagcgctc tcgcgttgca
     5101 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt
     5161 gcatttttgt tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg
     5221 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     5281 gcgttgcatt tttgttctac aaaatgaagc acagatgctt cgttaacaaa gatatgctat
     5341 tgaagtgcaa gatggaaacg cagaaaatga accggggatg cgacgtgcaa gattacctat
     5401 gcaatagatg caatagtttc tccaggaacc gaaatacata cattgtcttc cgtaaagcgc
     5461 tagactatat attattatac aggttcaaat atactatctg tttcagggaa aactcccagg
     5521 ttcggatgtt caaaattcaa tgatgggtaa caagtacgat cgtaaatctg taaaacagtt
     5581 tgtcggatat taggctgtat ctcctcaaag cgtattcgaa tatcattgag aagctgcagg
     5641 caagtgcaca aacaatactt aaataaatac tactcagtaa taacctattt cttagcattt
     5701 ttgacgaaat ttgctatttt gttagagtct tttacaccat ttgtctccac acctccgctt
     5761 acatcaacac caataacgcc atttaatcta agcgcatcac caacattttc tggcgtcagt
     5821 ccaccagcta acataaaatg taagctttcg gggctctctt gccttccaac ccagtcagaa
     5881 atcgagttcc aatccaaaag ttcacctgtc ccacctgctt ctgaatcaaa caagggaata
     5941 aacgaatgag gtttctgtga agctgcactg agtagtatgt tgcagtcttt tggaaatacg
     6001 agtcttttaa taactggcaa accgaggaac tcttggtatt cttgccacga ctcatctcca
     6061 tgcagttgga cgatatcaat gccgtaatca ttgaccagag ccaaaacatc ctccttaggt
     6121 tgattacgaa acacgccaac caagtatttc ggagtgcctg aactattttt atatgctttt
     6181 acaagacttg aaattttcct tgcaataacc gggtcaattg ttctctttct attgggcaca
     6241 catataatac ccagcaagtc agcatcggaa tctagagcac attctgcggc ctctgtgctc
     6301 tgcaagccgc aaactttcac caatggacca gaactacctg tgaaattaat aacagacata
     6361 ctccaagctg cctttgtgtg cttaatcacg tatactcacg tgctcaatag tcaccaatgc
     6421 cctccctctt ggccctctcc ttttcttttt tcgaccgaat ttcttgaaga cgaaagggcc
     6481 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     6541 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6601 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6661 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6721 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6781 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6841 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6901 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6961 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7021 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7081 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7141 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7201 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7261 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7321 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7381 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7441 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7501 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7561 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7621 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7681 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7741 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7801 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     7861 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7921 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7981 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8041 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8101 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8161 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8221 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8281 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8341 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     8401 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     8461 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     8521 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     8581 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     8641 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga
     8701 attaattcga gctcggta
//
