LOCUS       40924_45628        6711 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway vector pUGW42 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6711)
  AUTHORS   Tanaka Y, Nakagawa T.
  TITLE     Gateway Vectors for transient expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6711)
  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 6711)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6711)
  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            "Unpublished"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..6711
                     /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        745..1090
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1111..1827
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    1837..1961
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1998..2028
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2082..2738
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3083..3385
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3429..3553)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3579..3831
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3840..3856)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4069..4524
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4806..4910
                     /label=AmpR promoter
     CDS             4911..5768
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5942..6530
                     /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 ggtccccaga ttagcctttt caatttcaga aagaatgcta acccacagat
      301 ggttagagag gcttacgcag caggtctcat caagacgatc tacccgagca ataatctcca
      361 ggaaatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg
      421 catcaagaac acagagaaag atatatttct caagatcaga agtactattc cagtatggac
      481 gattcaaggc ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt
      541 agttcccact gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact
      601 cgccgtaaag actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa
      661 aatcttcgtc aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
      721 agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
      781 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
      841 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
      901 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
      961 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     1021 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     1081 ggagagaaca cgggggactc tagaatgagc atggtgagca agggcgagga gctgttcacc
     1141 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1201 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1261 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag
     1321 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1381 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1441 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1501 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1561 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1621 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1681 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     1741 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1801 actctcggca tggacgagct gtacaaggct gttatcacaa gtttgtacaa aaaagctgaa
     1861 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag
     1921 actacataat actgtaaaac acaacatatc cagtcactat ggcggcccct cgaggggccg
     1981 cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga
     2041 gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     2101 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     2161 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     2221 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     2281 tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     2341 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     2401 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     2461 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     2521 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     2581 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     2641 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     2701 aattacaaca gtactgcgat gagtggcagg gcggggcgta aacgcgtgga tccggcttac
     2761 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata
     2821 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt
     2881 gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt
     2941 ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg
     3001 gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct
     3061 gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg
     3121 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt
     3181 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     3241 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     3301 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     3361 cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtctgc
     3421 aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
     3481 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct
     3541 tgtacaaagt ggtgataacc tagagctcga atttccccga tcgttcaaac atttggcaat
     3601 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     3661 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     3721 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc
     3781 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattca
     3841 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3901 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3961 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     4021 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4081 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4141 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4201 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4261 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4321 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4381 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     4441 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4501 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4561 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4621 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4681 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4741 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4801 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4861 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4921 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4981 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5041 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5101 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5161 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5221 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5281 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5341 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5401 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5461 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5521 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5581 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5641 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5701 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5761 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5821 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5881 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6061 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6121 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6181 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6241 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6301 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6361 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6421 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6481 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6541 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6601 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6661 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
