LOCUS       40924_1254        6531 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GWluc-basic, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6531)
  AUTHORS   Del Vecchio I, Zuccotti A, Canneva F, Lenzken SC, Racchi M.
  TITLE     Development of the first Gateway firefly luciferase vector and use 
            of reverse transcriptase in FLOE (Fluorescently Labeled 
            Oligonucleotide Extension) reactions
  JOURNAL   Plasmid 58 (3), 269-274 (2007)
  PUBMED    17707908
REFERENCE   2  (bases 1 to 6531)
  AUTHORS   Del Vecchio I, Zuccotti A, Canneva F, Lanni C, Lenzken C, Govoni S, 
            Racchi M.
  TITLE     Improved firefly luciferase vector to rapidly recombine promoter 
            regions
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6531)
  AUTHORS   Del Vecchio I, Zuccotti A, Canneva F, Lanni C, Lenzken C, Govoni S, 
            Racchi M.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-FEB-2007) Centre of Excellence in Applied Biology - 
            Experimental and Applied Pharmacology Dept., University of Pavia, 
            Via Taramelli 14, Pavia 27100, Italy
REFERENCE   4  (bases 1 to 6531)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6531)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2007"; volume: "58"; issue: "3"; pages: "269-274"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (22-FEB-2007) Centre of Excellence in Applied Biology - Experimental
            and Applied Pharmacology Dept., University of Pavia, Via Taramelli 
            14, Pavia 27100, Italy"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..28
                     /label=multiple cloning site 1
                     /note="multiple cloning site 1"
     protein_bind    33..157
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        182..212
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             266..922
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1267..1569
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1613..1737)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    1742..1770
                     /label=multiple cloning site 2
                     /note="multiple cloning site 2"
     CDS             1801..3450
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    complement(3494..3615)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3717..3728
                     /label=multiple cloning site 3
                     /note="multiple cloning site 3"
     rep_origin      4031
                     /label=ColE1-derived plasmid replication origin
                     /note="ColE1-derived plasmid replication origin"
     rep_origin      complement(4034..4622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4796..5653)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5654..5758)
                     /label=AmpR promoter
     rep_origin      5785..6240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    6371..6419
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    6433..6524
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccat caacaagttt gtacaaaaaa gctgaacgag
       61 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
      121 cataatactg taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg
      181 ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag
      241 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
      301 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
      361 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
      421 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
      481 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
      541 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
      601 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
      661 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
      721 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
      781 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
      841 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
      901 cgatgagtgg cagggcgggg cgtaaagatc tggatccggc ttactaaaag ccagataaca
      961 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
     1021 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
     1081 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
     1141 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
     1201 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
     1261 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     1321 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     1381 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     1441 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     1501 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     1561 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga
     1621 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
     1681 tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggttga
     1741 tgggctcgag atctgcgatc taagtaagct tggcattccg gtactgttgg taaagccacc
     1801 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga
     1861 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
     1921 gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc
     1981 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
     2041 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
     2101 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt
     2161 tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
     2221 aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga
     2281 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
     2341 tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga
     2401 tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg
     2461 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
     2521 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
     2581 cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac
     2641 aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg
     2701 attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct
     2761 aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat
     2821 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     2881 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     2941 acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt
     3001 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     3061 ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct
     3121 ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa
     3181 caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt
     3241 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     3301 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac
     3361 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     3421 aaggccaaga agggcggaaa gatcgccgtg taattctaga gtcggggcgg ccggccgctt
     3481 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3541 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3601 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3661 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     3721 ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3781 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3841 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3901 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3961 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4261 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4321 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4381 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4441 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4501 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4561 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4621 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4681 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4741 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4801 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4861 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4921 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4981 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5041 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5101 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5161 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5221 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5281 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5341 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5401 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5461 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5521 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5581 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5641 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5701 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5761 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5821 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5881 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5941 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6001 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6061 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6121 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6181 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6241 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     6301 tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg
     6361 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     6421 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
     6481 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat a
//
