LOCUS       40924_45618        5858 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW4 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5858)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5858)
  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 5858)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5858)
  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            "Unpublished"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..5858
                     /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    261..385
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        422..452
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             506..1162
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1507..1809
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1853..1977)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1982..2698
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     terminator      2726..2978
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2987..3003)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3216..3671
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3953..4057
                     /label=AmpR promoter
     CDS             4058..4915
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5089..5677
                     /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 ggctctaatc acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg
      301 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
      361 aaacacaaca tatccagtca ctatggcggc ccctcgaggg gccgcattag gcaccccagg
      421 ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag
      481 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
      541 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
      601 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
      661 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
      721 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
      781 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
      841 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
      901 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
      961 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1021 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1081 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1141 cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca
     1201 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
     1261 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
     1321 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
     1381 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
     1441 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
     1501 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     1561 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     1621 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     1681 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     1741 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     1801 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga
     1861 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
     1921 tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtgat
     1981 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2041 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2101 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2161 cctcgtgacc accttcacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2221 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2281 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2341 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2401 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2461 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2521 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2581 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2641 cctgctggag ttcgtgaccg ccgccgggat cactcacggc atggacgagc tgtacaagta
     2701 aagcggcccg agctcgaatt tccccgatcg ttcaaacatt tggcaataaa gtttcttaag
     2761 attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa
     2821 gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag
     2881 agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga
     2941 taaattatcg cgcgcggtgt catctatgtt actagatcgg gaattcactg gccgtcgttt
     3001 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3061 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3121 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     3181 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3241 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3301 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3361 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3421 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3481 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3541 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     3601 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3661 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3721 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3781 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3841 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3901 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3961 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4021 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4081 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4141 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4201 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4261 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4321 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4381 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4441 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4501 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4561 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4621 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4681 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4741 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4801 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4861 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5221 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5701 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5761 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5821 gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
