LOCUS       40924_18486        5789 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pETAT-12, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5789)
  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 5789)
  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 5789)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5789)
  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..5789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..38
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    319..416
                     /note="multiple cloning site; MCS"
     misc_feature    complement(378..410)
                     /note="Region: synthetic HIV-1 TAT protein transduction
                     domain (PTD)"
     CDS             complement(417..434)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(444..461)
                     /label=6xHis
                     /note="6xHis affinity tag"
     RBS             complement(480..502)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(517..541)
                     /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(542..560)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        869..946
                     /label=lacI promoter
     CDS             947..2026
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2042..2063
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3341..3529
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3634..3776
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3962..4550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4724..5581)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5582..5686)
                     /label=AmpR promoter
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcctcgagt gcggccgcaa gcttgtcgac ggagctcgaa
      361 ttcggatccg aggtaccagc acgagcctga cgagcagcag cacgagcgta catatggctg
      421 ccgcgcggca ccaggccgct gctgtgatga tgatgatgat ggctgctgcc catggtatat
      481 ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc
      541 ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc
      601 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
      661 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
      721 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
      781 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
      841 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
      901 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
      961 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     1021 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     1081 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     1141 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     1201 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     1261 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     1321 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     1381 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     1441 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     1501 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     1561 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     1621 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     1681 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     1741 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     1801 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     1861 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     1921 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     1981 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2041 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
     2101 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2161 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
     2221 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     2281 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     2341 agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg
     2401 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga
     2461 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc
     2521 aaggatcgct cgcggctctt accagcctaa cttcgatcac tggaccgctg atcgtcacgg
     2581 cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc
     2641 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
     2701 gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca
     2761 attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc
     2821 cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt
     2881 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     2941 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     3001 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
     3061 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
     3121 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
     3181 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
     3241 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
     3301 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     3361 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     3421 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     3481 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3541 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3601 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     3661 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3721 catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3781 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3841 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3901 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3961 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4021 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4081 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4141 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4201 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4261 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4321 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4381 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4441 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4501 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4561 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4621 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4681 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4741 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4801 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4861 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4921 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4981 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     5041 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5101 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5161 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5221 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5281 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     5341 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5401 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5461 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5521 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5581 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5641 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5701 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5761 gtatcacgag gccctttcgt cttcaagaa
//
