LOCUS       40924_33003        5418 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNB-3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  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 5418)
  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 5418)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5418)
  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..5418
                     /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"
     RBS             complement(222..244)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(259..283)
                     /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(284..302)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        611..688
                     /label=lacI promoter
     CDS             689..1768
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1784..1805
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2580..2768
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2873..3015
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3201..3789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3963..4820)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4821..4925)
                     /label=AmpR promoter
     rep_origin      complement(4952..5407)
                     /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 ccatatgtat atctccttct taaagttaaa
      241 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
      301 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
      361 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
      421 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
      481 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
      541 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
      601 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
      661 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
      721 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
      781 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
      841 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
      901 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
      961 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     1021 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     1081 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     1141 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     1201 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     1261 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     1321 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     1381 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     1441 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     1501 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     1561 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     1621 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     1681 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     1741 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     1801 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     1861 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     1921 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     1981 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     2041 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     2101 gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct
     2161 ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg
     2221 actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg
     2281 tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca
     2341 tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc
     2401 attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct
     2461 cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct
     2521 ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag
     2581 tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa
     2641 cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc
     2701 ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg
     2761 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     2821 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     2881 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     2941 gattgtactg agagtgcacc atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     3001 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3061 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3601 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4261 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4921 ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa acgttaatat tttgttaaaa
     4981 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5041 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5101 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5161 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5221 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5281 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5341 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5401 ggcgcgtccc attcgcca
//
