LOCUS       40924_45613        6789 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW35 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6789)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6789)
  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 6789)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6789)
  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            "Unpublished"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..6789
                     /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..3632
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     terminator      3657..3909
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3918..3934)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4147..4602
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4884..4988
                     /label=AmpR promoter
     CDS             4989..5846
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6020..6608
                     /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 gcatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg
     2041 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa
     2101 ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt
     2161 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg
     2221 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag
     2281 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca
     2341 tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc
     2401 aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg
     2461 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg
     2521 attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg
     2581 gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct
     2641 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg
     2701 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat
     2761 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt
     2821 acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc
     2881 tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct
     2941 ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat
     3001 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg
     3061 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga
     3121 aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg
     3181 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt
     3241 ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt
     3301 ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc
     3361 aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac
     3421 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg
     3481 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg
     3541 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca
     3601 taaaggccaa gaagggcgga aagatcgccg tgtaagctta gagctcgaat ttccccgatc
     3661 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga
     3721 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     3781 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     3841 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
     3901 tactagatcg ggaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3961 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4021 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     4081 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4141 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4201 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4261 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4321 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4381 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4441 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     4501 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4561 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4621 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4681 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4741 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4801 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4861 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4921 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4981 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5041 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5101 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5161 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5221 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5281 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5341 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5401 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5461 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5521 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5581 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5641 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5701 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5761 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5821 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5881 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5941 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6001 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6061 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     6181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6721 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6781 aagcggaag
//
