LOCUS       40924_5719        6160 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector for the regulated expression of 
            N-terminally tagged HP-thioredoxin fusion proteins in bacterial 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6160)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6160)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     RBS             312..334
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             346..675
                     /codon_start=1
                     /label=HP-thioredoxin
                     /note="His-Patch thioredoxin"
                     /translation="MGSDKIIHLTDDSFDTDVLKADGAILVDFWAHWCGPCKMIAPILD
                     EIADEYQGKLTVAKLNIDHNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEF
                     LDANLA"
     CDS             691..705
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     protein_bind    718..842
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        867..897
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             951..1607
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1952..2254
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2298..2422)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2436..2477
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             2487..2504
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2730..2816
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2908..2935
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2955..3046
                     /label=AmpR promoter
     CDS             3047..3904
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4078..4666
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4852..4992)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5259..6134)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagaaataa ttttgtttaa ctttaagaag gagatataca tacccatggg atctgataaa
      361 attattcatc tgactgatga ttcttttgat actgatgtac ttaaggcaga tggtgcaatc
      421 ctggttgatt tctgggcaca ctggtgcggt ccgtgcaaaa tgatcgctcc gattctggat
      481 gaaatcgctg acgaatatca gggcaaactg accgttgcaa aactgaacat cgatcacaac
      541 ccgggcactg cgccgaaata tggcatccgt ggtatcccga ctctgctgct gttcaaaaac
      601 ggtgaagtgg cggcaaccaa agtgggtgca ctgtctaaag gtcagttgaa agagttcctc
      661 gacgctaacc tggccggctc tggatccggt gatgacgatg acaagctggg aattatcaca
      721 agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat
      781 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
      841 tggcggccgc attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt
      901 ggattttgag ttaggatccg gcgagatttt caggagctaa ggaagctaaa atggagaaaa
      961 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     1021 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     1081 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     1141 cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga
     1201 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     1261 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     1321 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     1381 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     1441 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     1501 tgatgccgct ggcgattcag gttcatcatg ccgtctgtga tggcttccat gtcggcagaa
     1561 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa acgcgtggat
     1621 ccggcttact aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa
     1681 gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtgtgcta tgaagcagcg
     1741 tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat
     1801 atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc
     1861 tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc
     1921 tcttttgctg acgagaacag ggactggtga aatgcagttt aaggtttaca cctataaaag
     1981 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
     2041 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact
     2101 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     2161 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     2221 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccg ttatacacag
     2281 ccagtctgca ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct
     2341 gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt
     2401 cagctttctt gtacaaagtg gtgatcaagc ttgaaggtaa gcctatccct aaccctctcc
     2461 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacggtct
     2521 ccagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     2581 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     2641 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     2701 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2761 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     2821 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     2881 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     2941 ttctacaaac tctttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3001 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3061 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3121 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3181 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3241 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     3301 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3361 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3421 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3481 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3541 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3601 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3661 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3721 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3781 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3841 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3901 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3961 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4021 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4081 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4141 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4201 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4261 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4321 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4381 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4441 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4501 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4561 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4621 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4681 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4741 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4801 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4861 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     4921 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     4981 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5041 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     5101 ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag
     5161 cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
     5221 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
     5281 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
     5341 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
     5401 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
     5461 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
     5521 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
     5581 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
     5641 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
     5701 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
     5761 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
     5821 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
     5881 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
     5941 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
     6001 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
     6061 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
     6121 tgcgcttcag ccatactttt catactcccg ccattcagag
//
