LOCUS       40924_14455        7072 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDEST375, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7072)
  AUTHORS   Giuraniuc CV, Macpherson M, Saka Y.
  TITLE     Gateway Vectors for Efficient Artificial Gene Assembly In Vitro and 
            Expression in Yeast Saccharomyces cerevisiae
  JOURNAL   PLoS ONE 8 (5), E64419 (2013)
  PUBMED    23675537
REFERENCE   2  (bases 1 to 7072)
  AUTHORS   Giuraniuc C, Macpherson M, Saka Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-FEB-2013) School of Medical Sciences, Institute of 
            Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen 
            AB25 2ZD, UK
REFERENCE   3  (bases 1 to 7072)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7072)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2013"; volume: "8"; issue: "5"; pages: "E64419"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-FEB-2013) School of Medical Sciences, Institute of Medical 
            Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, 
            UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(5..351)
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             complement(1187..1987)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        1990..2204
                     /label=URA3 promoter
     promoter        2301..2405
                     /label=AmpR promoter
     CDS             2406..3263
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3437..4025
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4265..4866)
                     /label=3' flanking MET15 YIp-In fragment
                     /note="3' flanking MET15 YIp-In fragment"
     terminator      complement(4920..5117)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    5143..5267
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5292..5322
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             5376..6032
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             6377..6679
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(6723..6847)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6897..6916)
                     /label=T7 reverse primer
                     /note="T7 reverse primer"
     terminator      6914..6961
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 caggtggatg ggggtaatag aattgtatct atgtatctga cgaccctgta ttacacgaag
       61 gaaatagtag atgaaaagac aagagagcaa gaaaaaggaa aaacttcctt tctaacagac
      121 gctttactta accttatata tatcttattt ttttcatcga gcgtgttcaa ttggacaagg
      181 tgccaccttt tcgacacttc ggttattatg ttacacagtt ttcatgagga tggcgccttg
      241 actaacttaa ttagtcattt gccaaccacc acagttcccc aatatcgaca gcttcacgtg
      301 ccatttgcca ttttgcgatc atgtgacctc aagagaaaaa gcaaaaaata atgagagcta
      361 agggattcga acccttgcat ccgaagatat cagagatttt agaggttaaa tccacctaaa
      421 tctgacgcct taaaccactc ggccaaactc ccattgtatg ttggaagaat tggatttaaa
      481 tgtctattat agcattaaat atctttaaat acattcaaga aatggcgaaa ttatttctta
      541 gtgttggaat aaaaatcaac tatcatctac taactagtat ttacgttact agtatattat
      601 catatacggt gttagaagat gacgcaaatg atgagaaata gtcatctaaa ttagtggaag
      661 ctgaaacgca aggattgata atgtaatagg atcaatgaat attaacatat aaaacgatga
      721 taataatatt tatagaattg tgtagaattg cagattccct tttatggatt cctaaatcct
      781 cgacgcctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      841 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      901 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
      961 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     1021 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     1081 accgaaacgc gcgagacgaa agggcgggta ataactgata taattaaatt gaagctctaa
     1141 tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc
     1201 atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat
     1261 cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca
     1321 catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac
     1381 cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa
     1441 taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt
     1501 ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt
     1561 cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac
     1621 cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca
     1681 atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact
     1741 tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat
     1801 ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt
     1861 ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat
     1921 taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt
     1981 tcgacatgat tcaattcatc attttttttt tattcttttt tttgatttcg gtttctttga
     2041 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     2101 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     2161 acccaactgc acagaacaaa aacatgcagg aaacgaagat aaattgaaaa gctggcctcg
     2221 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2281 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2341 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2401 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2461 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2521 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2581 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2641 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2701 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2761 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2821 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2881 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2941 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3001 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3061 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3121 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3181 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3241 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3301 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3361 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3421 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3481 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3541 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3601 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3661 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3721 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3781 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3841 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3901 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3961 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4021 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4081 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4141 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4201 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4261 ggcagatccg tatggaaagt agcatccaac caaatcccgc gaagttttcc tgatttacat
     4321 aacaagcaaa tactaaaaac gttaacacac atgtgcttct ttattgtaaa gaaacctcca
     4381 tcatcctctt ttgtaacttg gtcctacaat aaatttatcc agtgtgacag ctttatagga
     4441 ggcgtaaagt agtctcatga agtagatgag tcttgttcac catatttttc ttctcgactg
     4501 cgaattaaca ctgttctttg atgttagaac aatttaggtt caaagtacga gtcacgacat
     4561 gtaacaaggg aaaaaaagga tattcatttc aataaagttc gttttataaa agtatagtac
     4621 ttgtgagaga aagtaggttt atacataatt ttacaactca ttacgcacac tcatggtttt
     4681 tggccagcga aaacagtttc aaaagattgc tggaagtctg caataatgtc atcaataaat
     4741 tcgataccaa cagagacacg aattaagtcc ttggtaacac cagatgccaa cttttctttg
     4801 tcatttaatt gtttgtgggt agtgaagtat ggagcaatga ctaaggtctt ggcatcacca
     4861 accctggaat tcgagctcgt tttcgacact ggatggcggc gttagtatcg aatcgacagc
     4921 agtatagcga ccagcattca catacgattg acgcatgata ttactttctg cgcacttaac
     4981 ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct aacatttgat
     5041 taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt gaaaacaaga
     5101 atctttttat tgtcagtact gagcggccgc tctagaacta ggacaagttt gtacaaaaaa
     5161 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     5221 aaacagacta cataatactg taaaacacaa catatccagt cactatggcg gccgcattag
     5281 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg
     5341 atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
     5401 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     5461 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     5521 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     5581 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     5641 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     5701 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     5761 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     5821 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     5881 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     5941 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     6001 aacagtactg cgatgagtgg cagggcgggg cgtaaagatc tggatccggc ttactaaaag
     6061 ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata
     6121 tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt
     6181 tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta
     6241 agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat
     6301 caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag
     6361 aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     6421 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc
     6481 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     6541 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     6601 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     6661 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg
     6721 accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
     6781 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca
     6841 aagtggttga ttcgaggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     6901 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     6961 gctgaaagga ggaactatat ccggattcga gggggggccc ggtaccccgg ggatcctcta
     7021 gagtcgacct gcaggcatgc atcgataagc ttagatctct cgaggcggcc gc
//
