LOCUS       40924_45603        7031 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW3 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7031)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7031)
  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 7031)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7031)
  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            "Unpublished"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..7031
                     /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    267..391
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        428..458
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             512..1189
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             1512..1814
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1858..1982)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2017..3825
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     terminator      3899..4151
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4160..4176)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4389..4844
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5126..5230
                     /label=AmpR promoter
     CDS             5231..6088
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6262..6850
                     /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 ggtcgactct aatcaaacaa gtttgtacaa aaaagctgaa cgagaaacgt
      301 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
      361 actgtaaaac acaacatatc cagtcactat ggcggcccct cgaggggccg cattaggcac
      421 cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga gttaggatcc
      481 gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
      541 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
      601 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
      661 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
      721 tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
      781 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
      841 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
      901 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
      961 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     1021 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     1081 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     1141 gtactgcgat gagtggcagg cggggcgtaa tctagaggat ccggcttact aaaagccaga
     1201 taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat
     1261 acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca
     1321 gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac
     1381 aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga
     1441 agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag
     1501 gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
     1561 ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg
     1621 ccagtgcacg tctgctgtca gataaagtcc cccgtgaact ttacccggtg gtgcatatcg
     1681 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg
     1741 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
     1801 tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat
     1861 agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa
     1921 tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg
     1981 gttcgatcta gaggatcccc gggtggtcag tcccttatgt tacgtcctgt agaaacccca
     2041 acccgtgaaa tcaaaaaact cgacggcctg tgggcattca gtctggatcg cgaaaactgt
     2101 ggaattgatc agcgttggtg ggaaagcgcg ttacaagaaa gccgggcaat tgctgtgcca
     2161 ggcagtttta acgatcagtt cgccgatgca gatattcgta attatgcggg caacgtctgg
     2221 tatcagcgcg aagtctttat accgaaaggt tgggcaggcc agcgtatcgt gctgcgtttc
     2281 gatgcggtca ctcattacgg caaagtgtgg gtcaataatc aggaagtgat ggagcatcag
     2341 ggcggctata cgccatttga agccgatgtc acgccgtatg ttattgccgg gaaaagtgta
     2401 cgtatcaccg tttgtgtgaa caacgaactg aactggcaga ctatcccgcc gggaatggtg
     2461 attaccgacg aaaacggcaa gaaaaagcag tcttacttcc atgatttctt taactatgcc
     2521 ggaatccatc gcagcgtaat gctctacacc acgccgaaca cctgggtgga cgatatcacc
     2581 gtggtgacgc atgtcgcgca agactgtaac cacgcgtctg ttgactggca ggtggtggcc
     2641 aatggtgatg tcagcgttga actgcgtgat gcggatcaac aggtggttgc aactggacaa
     2701 ggcactagcg ggactttgca agtggtgaat ccgcacctct ggcaaccggg tgaaggttat
     2761 ctctatgaac tgtgcgtcac agccaaaagc cagacagagt gtgatatcta cccgcttcgc
     2821 gtcggcatcc ggtcagtggc agtgaagggc gaacagttcc tgattaacca caaaccgttc
     2881 tactttactg gctttggtcg tcatgaagat gcggacttgc gtggcaaagg attcgataac
     2941 gtgctgatgg tgcacgacca cgcattaatg gactggattg gggccaactc ctaccgtacc
     3001 tcgcattacc cttacgctga agagatgctc gactgggcag atgaacatgg catcgtggtg
     3061 attgatgaaa ctgctgctgt cggctttaac ctctctttag gcattggttt cgaagcgggc
     3121 aacaagccga aagaactgta cagcgaagag gcagtcaacg gggaaactca gcaagcgcac
     3181 ttacaggcga ttaaagagct gatagcgcgt gacaaaaacc acccaagcgt ggtgatgtgg
     3241 agtattgcca acgaaccgga tacccgtccg caaggtgcac gggaatattt cgcgccactg
     3301 gcggaagcaa cgcgtaaact cgacccgacg cgtccgatca cctgcgtcaa tgtaatgttc
     3361 tgcgacgctc acaccgatac catcagcgat ctctttgatg tgctgtgcct gaaccgttat
     3421 tacggatggt atgtccaaag cggcgatttg gaaacggcag agaaggtact ggaaaaagaa
     3481 cttctggcct ggcaggagaa actgcatcag ccgattatca tcaccgaata cggcgtggat
     3541 acgttagccg ggctgcactc aatgtacacc gacatgtgga gtgaagagta tcagtgtgca
     3601 tggctggata tgtatcaccg cgtctttgat cgcgtcagcg ccgtcgtcgg tgaacaggta
     3661 tggaatttcg ccgattttgc gacctcgcaa ggcatattgc gcgttggcgg taacaagaaa
     3721 gggatcttca ctcgcgaccg caaaccgaag tcggcggctt ttctgctgca aaaacgctgg
     3781 actggcatga acttcggtga aaaaccgcag cagggaggca aacaatgaat caacaactct
     3841 cctggcgcac catcgtcggc tacagcctcg ggaattgcta ccgagctcga atttccccga
     3901 tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat
     3961 gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat
     4021 gacgttattt atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc
     4081 gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat
     4141 gttactagat cgggaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     4201 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4261 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     4321 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4381 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4441 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4501 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4561 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4621 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4681 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     4741 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4801 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4861 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4921 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4981 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5041 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5101 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5161 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5221 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5281 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5341 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5401 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5461 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5521 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5581 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5641 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5701 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5761 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5821 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5881 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5941 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6001 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6061 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6121 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6181 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6241 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6301 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6361 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     6421 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6481 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6541 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6601 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6661 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6721 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6781 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6841 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6901 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6961 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7021 ggaagcggaa g
//
