LOCUS       pT-REx-DEST31.        7559 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) destination vector enabling N-terminal 6xHis-tagging and 
            tetracycline-inducible high-level expression.
ACCESSION   .
VERSION     .
KEYWORDS    pT-REx-DEST31
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7559)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7559)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        313..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    518..536
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    539..557
                     /gene="tetO"
                     /label=tetracycline repressor TetR binding site
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             661..663
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             673..690
                     /label=6xHis
                     /note="6xHis affinity tag"
     protein_bind    715..839
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        864..894
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             948..1604
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1949..2251
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2295..2419)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2480..2498)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2509..2525)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2930..3064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3190..3645
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3764..4121
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4172..4963
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5030..5078
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        5384..5488
                     /label=AmpR promoter
     CDS             5489..6346
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6520..7108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgcatgtcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
       61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga tagagatctc
      541 cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat cgcctggaga
      601 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggacc
      661 atggcgtact accatcacca tcaccatcac accggtgata tcctcgagcc catcacaagt
      721 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
      781 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
      841 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
      901 ttttgagtta ggatccggcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
      961 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1021 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1081 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1141 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     1201 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1261 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1321 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1381 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1441 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1501 tgccgctggc gattcaggtt catcatgccg tctgtgatgg cttccatgtc ggcagaatgc
     1561 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg
     1621 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     1681 tatatactga tatgtatacc cgaagtatgt caaaaagagg tgtgctatga agcagcgtat
     1741 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     1801 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     1861 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     1921 tttgctgacg agaacaggga ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     1981 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     2041 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     2101 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     2161 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     2221 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccgtta tacacagcca
     2281 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     2341 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     2401 ctttcttgta caaagtggtg atgggcggcc gctctagagg gcccaagctt acgcgtgcat
     2461 gcgacgtcat agctctctcc ctatagtgag tcgtattata agctaggcac tggccgtcgt
     2521 tttacaacgt cgtgactggg aaaactgcta gcttgggatc tttgtgaagg aaccttactt
     2581 ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat
     2641 aaaattttta agtgtataat gtgttaaact agctgcatat gcttgctgct tgagagtttt
     2701 gcttactgag tatgatttat gaaaatatta tacacaggag ctagtgattc taattgtttg
     2761 tgtattttag attcacagtc ccaaggctca tttcaggccc ctcagtcctc acagtctgtt
     2821 catgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2881 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2941 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3001 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3061 gatcgatcct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattggct
     3121 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3181 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3541 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3601 aaatatttaa cgcgaatttt aacaaaatat taacgtttac aatttcgcct gatgcggtat
     3661 tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat
     3721 ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc
     3781 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3841 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3901 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3961 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4021 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4081 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca
     4141 caacagtctc gaacttaagg ctagagccac catgattgaa caagatggat tgcacgcagg
     4201 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4261 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4321 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4381 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4441 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4501 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4561 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4621 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4681 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4741 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4801 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4861 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4921 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4981 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc
     5041 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatccg
     5101 cgtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     5161 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5221 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5281 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5341 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5401 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5461 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5521 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5581 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5641 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5701 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5761 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5821 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5881 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5941 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6001 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6061 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6121 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6181 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6241 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6301 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6361 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6421 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6661 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6901 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7141 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7201 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     7261 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     7321 gcgttggccg attcattaat gcagagcttg caattcgcgc gtttttcaat attattgaag
     7381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7501 tattatcatg acattaacct ataaaaatag gcgtagtacg aggccctttc actcattag
//
