LOCUS       40924_18491        5451 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETAT-2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5451)
  AUTHORS   Yang Y, Ma J, Song Z, Wu M.
  TITLE     HIV-1 TAT-mediated protein transduction and subcellular localization
            using novel expression vectors
  JOURNAL   FEBS Lett. 532 (1-2), 36-44 (2002)
  PUBMED    12459459
REFERENCE   2  (bases 1 to 5451)
  AUTHORS   Yang YH, Wu M.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2002) Department of Molecular and Cellular 
            Biology, School of Life Sciences, University of Science and 
            Technology of China, 96 Jingzai Road, Hefei, Anhui 230026, P.R. 
            China
REFERENCE   3  (bases 1 to 5451)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5451)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "FEBS Lett. 
            532 (1-2), 36-44 (2002)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUN-2002) Department of Molecular and Cellular Biology, School 
            of Life Sciences, University of Science and Technology of China, 96 
            Jingzai Road, Hefei, Anhui 230026, P.R. China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    158..211
                     /note="multiple cloning site; MCS"
     misc_feature    complement(212..244)
                     /note="contains mutation; Region: synthetic HIV-1 TAT
                     protein transduction domain (PTD)"
     RBS             complement(255..277)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(292..316)
                     /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(317..335)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        644..721
                     /label=lacI promoter
     CDS             722..1801
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1817..1838
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2613..2801
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2906..3048
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3234..3822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3996..4853)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4854..4958)
                     /label=AmpR promoter
     rep_origin      complement(4985..5440)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgaggtac cagcacgagc ctgacgagca gcagcacgag
      241 cgtacatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt
      301 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc
      361 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
      421 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
      481 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
      541 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
      601 ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg
      661 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
      721 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
      781 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
      841 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
      901 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
      961 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     1021 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     1081 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     1141 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     1201 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     1261 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     1321 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     1381 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     1441 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     1501 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     1561 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     1621 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     1681 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     1741 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     1801 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga
     1861 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     1921 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     1981 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
     2041 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     2101 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga
     2161 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
     2221 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     2281 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     2341 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     2401 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
     2461 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
     2521 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     2581 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     2641 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     2701 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     2761 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2821 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2881 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     2941 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg
     3001 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     3061 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3661 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     4321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4981 cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5041 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5101 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5161 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5221 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5281 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5341 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5401 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc a
//
