LOCUS       40924_45578        6688 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW23 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6688)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6688)
  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 6688)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6688)
  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            "Unpublished"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..6688
                     /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        763..1108
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    1130..1254
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1291..1321
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1375..2031
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2376..2678
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2722..2846)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2857..3510
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             3517..3534
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     terminator      3556..3808
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3817..3833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4046..4501
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4783..4887
                     /label=AmpR promoter
     CDS             4888..5745
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5919..6507
                     /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 ggtctagacc caagcttgca tgcctgcagg tccccagatt agccttttca atttcagaaa
      301 gaatgctaac ccacagatgg ttagagaggc ttacgcagca ggtctcatca agacgatcta
      361 cccgagcaat aatctccagg aaatcaaata ccttcccaag aaggttaaag atgcagtcaa
      421 aagattcagg actaactgca tcaagaacac agagaaagat atatttctca agatcagaag
      481 tactattcca gtatggacga ttcaaggctt gcttcacaaa ccaaggcaag taatagagat
      541 tggagtctct aaaaaggtag ttcccactga atcaaaggcc atggagtcaa agattcaaat
      601 agaggaccta acagaactcg ccgtaaagac tggcgaacag ttcatacaga gtctcttacg
      661 actcaatgac aagaagaaaa tcttcgtcaa catggtggag cacgacacac ttgtctactc
      721 caaaaatatc aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag
      781 ggtaatatcc ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa
      841 gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat
      901 cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
      961 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
     1021 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
     1081 aggaagttca tttcatttgg agagaacacg ggggactcta gagttatcaa caagtttgta
     1141 caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt
     1201 gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac tatggcggcc
     1261 cctcgagggg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     1321 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
     1381 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     1441 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     1501 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1561 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     1621 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1681 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1741 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1801 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1861 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1921 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1981 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaagatct
     2041 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     2101 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     2161 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     2221 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     2281 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     2341 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     2401 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     2461 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     2521 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     2581 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     2641 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     2701 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     2761 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     2821 cgttcagctt tcttgtacaa agtggttgat aacagcatgt cccctatact aggttattgg
     2881 aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat
     2941 gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg
     3001 ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct
     3061 atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag
     3121 cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga
     3181 attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa
     3241 atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta
     3301 acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg
     3361 tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa
     3421 attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc
     3481 acgtttggtg gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg atccgcttag
     3541 agctcgaatt tccccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
     3601 gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata
     3661 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
     3721 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
     3781 cgcgcggtgt catctatgtt actagatcgg gaattcactg gccgtcgttt tacaacgtcg
     3841 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     3901 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3961 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4021 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4081 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4141 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4201 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4261 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4321 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4381 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4441 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4501 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4561 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4621 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4681 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4741 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4861 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4981 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5041 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5101 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5161 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5281 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5401 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5461 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5581 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5641 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5701 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5761 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5821 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6001 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6061 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6541 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6601 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6661 agcgagtcag tgagcgagga agcggaag
//
