LOCUS       40924_45608        6948 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW33 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6948)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6948)
  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 6948)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6948)
  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            "Unpublished"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..6948
                     /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"
     protein_bind    256..380
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        417..447
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             501..1157
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1502..1804
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1848..1972)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1983..3791
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     terminator      3816..4068
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4077..4093)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4306..4761
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5043..5147
                     /label=AmpR promoter
     CDS             5148..6005
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6179..6767
                     /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 ggtctagagt tatcaacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata
      301 aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca
      361 caacatatcc agtcactatg gcggcccctc gaggggccgc attaggcacc ccaggcttta
      421 cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt
      481 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
      541 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
      601 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
      661 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
      721 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
      781 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
      841 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
      901 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
      961 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     1021 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     1081 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     1141 agtggcaggg cggggcgtaa agatctggat ccggcttact aaaagccaga taacagtatg
     1201 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta
     1261 tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta
     1321 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag
     1381 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct
     1441 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga
     1501 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     1561 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg
     1621 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     1681 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     1741 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     1801 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga
     1861 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat
     1921 tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg gttgataaca
     1981 gcatgttacg tcctgtagaa accccaaccc gtgaaatcaa aaaactcgac ggcctgtggg
     2041 cattcagtct ggatcgcgaa aactgtggaa ttgatcagcg ttggtgggaa agcgcgttac
     2101 aagaaagccg ggcaattgct gtgccaggca gttttaacga tcagttcgcc gatgcagata
     2161 ttcgtaatta tgcgggcaac gtctggtatc agcgcgaagt ctttataccg aaaggttggg
     2221 caggccagcg tatcgtgctg cgtttcgatg cggtcactca ttacggcaaa gtgtgggtca
     2281 ataatcagga agtgatggag catcagggcg gctatacgcc atttgaagcc gatgtcacgc
     2341 cgtatgttat tgccgggaaa agtgtacgta tcaccgtttg tgtgaacaac gaactgaact
     2401 ggcagactat cccgccggga atggtgatta ccgacgaaaa cggcaagaaa aagcagtctt
     2461 acttccatga tttctttaac tatgccggaa tccatcgcag cgtaatgctc tacaccacgc
     2521 cgaacacctg ggtggacgat atcaccgtgg tgacgcatgt cgcgcaagac tgtaaccacg
     2581 cgtctgttga ctggcaggtg gtggccaatg gtgatgtcag cgttgaactg cgtgatgcgg
     2641 atcaacaggt ggttgcaact ggacaaggca ctagcgggac tttgcaagtg gtgaatccgc
     2701 acctctggca accgggtgaa ggttatctct atgaactgtg cgtcacagcc aaaagccaga
     2761 cagagtgtga tatctacccg cttcgcgtcg gcatccggtc agtggcagtg aagggcgaac
     2821 agttcctgat taaccacaaa ccgttctact ttactggctt tggtcgtcat gaagatgcgg
     2881 acttgcgtgg caaaggattc gataacgtgc tgatggtgca cgaccacgca ttaatggact
     2941 ggattggggc caactcctac cgtacctcgc attaccctta cgctgaagag atgctcgact
     3001 gggcagatga acatggcatc gtggtgattg atgaaactgc tgctgtcggc tttaacctct
     3061 ctttaggcat tggtttcgaa gcgggcaaca agccgaaaga actgtacagc gaagaggcag
     3121 tcaacgggga aactcagcaa gcgcacttac aggcgattaa agagctgata gcgcgtgaca
     3181 aaaaccaccc aagcgtggtg atgtggagta ttgccaacga accggatacc cgtccgcaag
     3241 gtgcacggga atatttcgcg ccactggcgg aagcaacgcg taaactcgac ccgacgcgtc
     3301 cgatcacctg cgtcaatgta atgttctgcg acgctcacac cgataccatc agcgatctct
     3361 ttgatgtgct gtgcctgaac cgttattacg gatggtatgt ccaaagcggc gatttggaaa
     3421 cggcagagaa ggtactggaa aaagaacttc tggcctggca ggagaaactg catcagccga
     3481 ttatcatcac cgaatacggc gtggatacgt tagccgggct gcactcaatg tacaccgaca
     3541 tgtggagtga agagtatcag tgtgcatggc tggatatgta tcaccgcgtc tttgatcgcg
     3601 tcagcgccgt cgtcggtgaa caggtatgga atttcgccga ttttgcgacc tcgcaaggca
     3661 tattgcgcgt tggcggtaac aagaaaggga tcttcactcg cgaccgcaaa ccgaagtcgg
     3721 cggcttttct gctgcaaaaa cgctggactg gcatgaactt cggtgaaaaa ccgcagcagg
     3781 gaggcaaaca atgagcttag agctcgaatt tccccgatcg ttcaaacatt tggcaataaa
     3841 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     3901 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     3961 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     4021 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg gaattcactg
     4081 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4141 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4201 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     4261 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     4321 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4381 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4441 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4501 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4561 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4621 ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga
     4681 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4741 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4801 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4861 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4921 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4981 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5041 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5101 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5161 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5221 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5281 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5341 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5401 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5461 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5521 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5581 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5641 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5701 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5761 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5821 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5881 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5941 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6001 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6061 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6121 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6181 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6241 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6301 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     6361 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6421 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6481 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6541 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6601 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6661 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6721 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6781 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6841 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6901 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
