LOCUS       V004232                 3710 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004232
VERSION     V004232
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 3710)
  AUTHORS   Ma Y.
  TITLE     Development of a dual-expression vector facilitated with
            selection-free PCR recombination cloning strategy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3710)
  AUTHORS   Ma Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-FEB-2017) Rongchang Campus, Southwest University, No.
            2 Tiansheng Road, Beibei District, Chongqing, Chongqing 400715,
            China
REFERENCE   3  (bases 1 to 3710)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3710)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (13-FEB-2017) Rongchang Campus, Southwest University, No. 2
            Tiansheng Road, Beibei District, Chongqing, Chongqing 400715, China"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        239..618
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        867..885
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    905..929
                     /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)."
     RBS             944..966
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    970..976
                     /label="Kozak sequence"
                     /note="Kozak sequence"
     CDS             985..1002
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             1264..1536
                     /gene="E"
                     /label="Lysis protein E"
                     /note="Lysis protein E from Enterobacteria phage phiX174.
                     Accession#: P03639"
     polyA_signal    1569..1793
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     terminator      1796..1843
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     rep_origin      complement(1958..2543)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2717..3574)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3575..3679)
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatcgatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc
      901 tagcggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
      961 aggagatata ccatgggcag cagccaccat catcaccacc acacctacca aacaatgccc
     1021 ccctgcaaaa aataaattca tataaaaaac atacagataa ccatctgcgg tgataaatta
     1081 tctctggcgg tgttgacata aataccactg gcggtgatac tgagcacatc agcaggacgc
     1141 actgaccacc atgaaggtga cgctcttaaa aattaagccc tgaagaaggg cagcattcaa
     1201 agcagaaggc tttggggtgt gtgatacgaa acgaagcatt ggttaaaaat taaggaggat
     1261 atcatggtac gctggacttt gtgggatacc ctcgctttcc tgctcctgtt gagtttattg
     1321 ctgccgtcat tgcttattat gttcatcccg tcaacattca aacggcctgt ctcatcatgg
     1381 aaggcgctga atttacggaa aacattatta atggcgtcga gcgtccggtt aaagccgctg
     1441 aattgttcgc gtttaccttg cgtgtacgcg caggaaacac tgacgttctt actgacgcag
     1501 aagaaaacgt gcgtcaaaaa ttacgtgcag aaggagtgat gtactgcagc caagcttctg
     1561 tgcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1621 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1681 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1741 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggctctagc
     1801 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgttcgaag gcggtaatac
     1861 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2461 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     2521 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2581 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2641 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2701 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2761 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2821 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2881 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2941 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3001 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3061 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3121 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3181 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3241 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3301 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3361 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3421 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3481 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3541 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3601 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3661 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
