LOCUS       40924_45548        6114 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW15 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6114)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6114)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2011) Contact:Tsuyoshi Nakagawa Shimane 
            University, Center for Integrated Research in Science; 1060 
            Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE   3  (bases 1 to 6114)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6114)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-APR-2011) Contact:Tsuyoshi Nakagawa Shimane University, Center 
            for Integrated Research in Science; 1060 Nishikawatsu, Matsue, 
            Shimane 690-8504, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     constructed using pUC119.
FEATURES             Location/Qualifiers
     source          1..6114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        745..1090
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1129..1218
                     /codon_start=1
                     /label=3xHA
                     /note="three tandem HA epitope tags"
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    1240..1364
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1401..1431
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1485..2141
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2486..2788
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2832..2956)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2982..3234
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3243..3259)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3472..3927
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4209..4313
                     /label=AmpR promoter
     CDS             4314..5171
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5345..5933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 catgcctgca ggtccccaga ttagcctttt caatttcaga aagaatgcta acccacagat
      301 ggttagagag gcttacgcag caggtctcat caagacgatc tacccgagca ataatctcca
      361 ggaaatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg
      421 catcaagaac acagagaaag atatatttct caagatcaga agtactattc cagtatggac
      481 gattcaaggc ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt
      541 agttcccact gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact
      601 cgccgtaaag actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa
      661 aatcttcgtc aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
      721 agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
      781 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
      841 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
      901 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
      961 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     1021 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     1081 ggagagaaca cgggggactc tagaatgagc gggttaatta acatctttta cccatacgat
     1141 gttcctgact atgcgggcta tccctatgac gtcccggact atgcaggatc ctatccatat
     1201 gacgttccag attacgctgc tcagtgcagc gctgttatca caagtttgta caaaaaagct
     1261 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa
     1321 cagactacat aatactgtaa aacacaacat atccagtcac tatggcggcc cctcgagggg
     1381 ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggattt
     1441 tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     1501 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     1561 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     1621 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     1681 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
     1741 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     1801 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     1861 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     1921 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     1981 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     2041 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     2101 atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt ggatccggct
     2161 tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt ataagaatat
     2221 atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc agcgtattac
     2281 agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt caatatctcc
     2341 ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa
     2401 gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa cggctctttt
     2461 gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata aaagagagag
     2521 ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat
     2581 ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc
     2641 ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc
     2701 ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa
     2761 cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac acagccagtc
     2821 tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt
     2881 tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcagctt
     2941 tcttgtacaa agtggtgata acctagagct cgaatttccc cgatcgttca aacatttggc
     3001 aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc
     3061 tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat
     3121 gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat
     3181 agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat
     3241 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3301 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3361 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     3421 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3481 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3541 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3601 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3661 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3721 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3781 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     3841 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3901 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3961 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4021 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4081 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4141 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4201 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4261 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4321 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4381 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4441 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4501 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4561 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4621 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4681 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4741 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4801 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4861 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4921 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4981 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5041 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5101 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5161 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5221 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5281 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5341 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5401 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5461 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5521 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5581 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5641 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5701 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5761 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5821 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5881 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5941 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6001 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6061 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
