LOCUS       40924_32450        5179 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for inducible high-level expression of
            C-terminally V5-6xHis-tagged proteins in Drosophila cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5179)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5179)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        401..827
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     protein_bind    853..977
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1002..1032
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1086..1742
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2087..2389
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2433..2557)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2571..2612
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             2622..2639
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    complement(2681..2815)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2959..2975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2983..2999)
                     /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)."
     promoter        complement(3007..3037)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3052..3073)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3361..3949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4123..4980)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4981..5085)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt aattcgttgc aggacaggat
      421 gtggtgcccg atgtgactag ctctttgctg caggccgtcc tatcctctgg ttccgataag
      481 agacccagaa ctccggcccc ccaccgccca ccgccacccc catacatatg tggtacgcaa
      541 gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag ctttgcatcc catacaagtc
      601 cccaaagtgg agaaccgaac caattcttcg cgggcagaac aaaagcttct gcacacgtct
      661 ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt gaatcgaacg aaagacccgt
      721 gtgtaaagcc gcgtttccaa aatgtataaa accgagagca tctggccaat gtgcatcagt
      781 tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa ctaaaggggg gatctagatc
      841 ggggtacatc aaacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
      901 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
      961 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     1021 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     1081 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1141 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1201 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1261 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     1321 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1381 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1441 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1501 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1561 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     1621 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct
     1681 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     1741 cgtaatctag aggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     1801 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggtg
     1861 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     1921 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     1981 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     2041 attgaaatga acggctcttt tgctgacgag aacagggact ggtgaaatgc agtttaaggt
     2101 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     2161 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     2221 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     2281 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     2341 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     2401 ctccgttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     2461 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     2521 tttacgtttc tcgttcagct ttcttgtaca aagtggttcg atggttcgaa ggtaagccta
     2581 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt
     2641 gagtttaaac ccgctgatca gcctcgactg tgccttctaa gatccagaca tgataagata
     2701 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2761 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2821 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     2881 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     2941 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3001 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3061 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3121 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3181 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3241 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3301 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3361 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3421 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3481 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3541 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3601 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3661 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3721 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3781 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3841 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3901 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3961 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4021 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4081 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4141 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4201 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4261 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4321 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4381 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4441 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4501 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4561 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4621 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4681 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4741 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4801 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4861 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4921 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4981 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5041 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5101 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5161 tatcacgagg ccctttcgt
//
