LOCUS       40924_18836        4403 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for cell-free expression and 
            C-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4403)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4403)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    98..222
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        247..277
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             331..987
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1332..1634
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1678..1802)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1822..1863
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             1873..1890
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1940..1987
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2156..2527
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     CDS             2551..3408
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3582..4167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga attatcaaca agtttgtaca aaaaagctga
      121 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
      181 gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc
      241 ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg
      301 gcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
      361 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
      421 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
      481 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
      541 ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
      601 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
      661 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
      721 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
      781 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
      841 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
      901 gttcatcatg ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
      961 tactgcgatg agtggcaggg cggggcgtaa agatctggat ccggcttact aaaagccaga
     1021 taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat
     1081 acccgaagta tgtcaaaaag aggtgtgcta tgaagcagcg tattacagtg acagttgaca
     1141 gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac
     1201 aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga
     1261 agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag
     1321 ggactggtga aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
     1381 ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg
     1441 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
     1501 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg
     1561 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
     1621 tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat
     1681 agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa
     1741 tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg
     1801 gttgataatt cgaagcttga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     1861 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa ctatatagaa taaaagaaga
     1921 aaccttagct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1981 ttttttgctg aaaggaggaa ctatatccgg attaacgctt acaatttagg tggcactttt
     2041 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2101 ccgctcatga gacaataacc ctgataaatg cttcaataat gtgaggaggg ccaccatggc
     2161 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     2221 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     2281 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     2341 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     2401 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     2461 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     2521 gcaggactga cacattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2581 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2641 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2701 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2761 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2821 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2881 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2941 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3001 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3061 ataccaaacg acgagagtga caccacgatg cctgtagcaa tgccaacaac gttgcgcaaa
     3121 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3181 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3241 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3301 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3361 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3421 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3481 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3541 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3601 cgcgtaatct gctgcttgca aacaaaaaaa cgcgctacca gcggtggttt gtttgccgga
     3661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3721 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4021 ggtaagcggc agggtcggaa caggagacgc acgagggagc ttccaggggg aaacgcctgg
     4081 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4141 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4201 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4261 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4321 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4381 cgttggccga ttcattaatg cag
//
