LOCUS       40924_14325        6422 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for inducible high-level expression of
            proteins in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6422)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6422)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    67..191
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        338..440
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             441..1097
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1442..1744
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1788..1912)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      1979..2026
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2392..2420)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        2533..2637
                     /label=AmpR promoter
     CDS             2638..3495
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3669..4257
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4443..4585)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4690..4878)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagatcacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
      121 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
      181 cagtcactat ggcggccgct aagttggcag catcacccga cgcactttgc gccgaataaa
      241 tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg
      301 ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta agaggttcca
      361 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
      421 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
      481 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
      541 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
      601 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
      661 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
      721 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
      781 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
      841 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
      901 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
      961 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     1021 ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     1081 agtggcaggg cggggcgtaa acgcgtggat ccggcttact aaaagccaga taacagtatg
     1141 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta
     1201 tgtcaaaaag aggtgtgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta
     1261 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag
     1321 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct
     1381 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag ggactggtga
     1441 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     1501 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg
     1561 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     1621 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     1681 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     1741 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga
     1801 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat
     1861 tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg gtgatgatcc
     1921 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1981 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     2041 tatatccgga tatccacagg acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc
     2101 caagtagcga agcgagcagg actgggcggc ggccaaagcg gtcggacagt gctccgagaa
     2161 cgggtgcgca tagaaattgc atcaacgcat atagcgctag cagcacgcca tagtgactgg
     2221 cgatgctgtc ggaatggacg atatcccgca agaggcccgg cagtaccggc ataaccaagc
     2281 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
     2341 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     2401 tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc
     2461 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2521 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2581 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2641 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2701 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2761 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2821 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2881 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2941 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3001 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3061 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3121 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3181 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3241 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3301 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3361 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3421 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3481 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3601 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3661 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3721 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3781 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4141 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4201 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4261 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4321 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4381 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4441 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt
     4501 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     4561 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     4621 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4681 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     4741 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     4801 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     4861 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     4921 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     4981 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     5041 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     5101 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     5161 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     5221 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     5281 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     5341 ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga
     5401 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     5461 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
     5521 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     5581 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
     5641 gtgacgatca gcggtccagt gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     5701 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     5761 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     5821 aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc
     5881 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     5941 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     6001 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     6061 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     6121 gctctcaagg gcatcggtcg atcgacgctc tcccttatgc gactcctgca ttaggaagca
     6181 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     6241 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     6301 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     6361 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     6421 cg
//
