LOCUS       V001996                 5660 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V001996
VERSION     V001996
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5660)
  AUTHORS   Boum Y, Myllykallio H.
  TITLE     Synthetic gene construct for expressing human thymidylate synthase
            in Escherichia coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5660)
  AUTHORS   Boum Y, Myllykallio H.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-FEB-2008) Institute of Genetics and Microbiology, U.
            Paris Sud, Bat. 409, Orsay 91405, France
REFERENCE   3  (bases 1 to 5660)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5660)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (22-FEB-2008) Institute of Genetics and Microbiology, U. Paris Sud,
            Bat. 409, Orsay 91405, France"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..54
                     /label="T5 promoter"
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    62..78
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             127..144
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             151..1089
                     /gene="TYMS"
                     /label="Thymidylate synthase"
                     /note="Thymidylate synthase from Homo sapiens. Accession#:
                     P04818"
     terminator      1117..1211
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             1255..1911
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     terminator      1979..2065
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(2124..2145)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2161..3240)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(3241..3318)
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    3510..3650
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3836..4424)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4598..5455)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5456..5560)
                     /label="AmpR promoter"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc atgccggtgg ccgggagcga actgccgcgt
      181 cgtccgttac cgccggcggc gcaggaacgt gatgcagaac cgcgcccgcc gcacggagaa
      241 ttgcagtacc tgggtcaaat tcagcatatc ctgcgttgtg gtgttcgcaa agatgatcgg
      301 acaggtaccg gcaccttaag cgtcttcggc atgcaggcga gatattcatt acgagatgaa
      361 tttccgctcc tgaccacgaa acgtgtcttt tggaaaggcg ttctggaaga gctcctgtgg
      421 tttatcaaag gctcaactaa cgccaaggaa ctgagcagca aaggcgtcaa aatttgggat
      481 gcgaacggca gccgcgattt tctggatagt ctgggcttca gtactcgtga ggaaggagac
      541 ctgggccctg tatatggttt ccaatggcgc cattttggtg ccgaataccg cgatatggaa
      601 agtgattact caggtcaggg agtagaccaa ctacagcgcg tgatcgatac cattaagacg
      661 aacccggatg atcgaagaat cataatgtgt gcttggaacc cgcgcgatct gccgctcatg
      721 gctcttccgc catgccatgc gctgtgtcaa ttttacgtag tcaacagtga actgtcgtgc
      781 caattgtacc agcgtagtgg cgacatgggc ttaggtgttc cgtttaacat cgcaagctat
      841 gctctgttga cctacatgat cgcacacatc accggcttaa aaccgggtga ttttatccac
      901 actctgggtg acgcccatat ttacctgaat cacatcgagc cactcaaaat tcagctgcag
      961 cgtgagccgc gcccgttccc gaaactgcgt attttgagaa aagtagagaa aatcgacgat
     1021 tttaaagctg aagatttcca gatcgaaggc tataatcctc acccgaccat taaaatggaa
     1081 atggctgtgt aatagaagct taattagctg agcttggact cctgttgata gatccagtaa
     1141 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     1201 ttggtgagaa tccaagctag cttggcgaga ttttcaggag ctaaggaagc taaaatggag
     1261 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     1321 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     1381 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1441 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
     1501 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1561 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1621 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1681 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1741 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1801 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1861 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt
     1921 taaggcagtt attggtgccc ttaaacgcct ggggtaatga ctctctagct tgaggcatca
     1981 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     2041 gaacgctctc ctgagtagga caaatccgcc ctctagatta cgtgcagtcg atgataagct
     2101 gtcaaacatg agaattgtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     2161 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2221 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     2281 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     2341 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     2401 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     2461 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     2521 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     2581 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     2641 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     2701 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     2761 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     2821 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     2881 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     2941 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     3001 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     3061 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     3121 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     3181 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     3241 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     3301 gcaccattcg atggtgtcgg aatttcgggc agcgttgggt cctggccacg ggtgcgcatg
     3361 atctagagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     3421 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     3481 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     3541 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     3601 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     3661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4261 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4561 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4621 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4681 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4741 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4801 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4861 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4921 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4981 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5041 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5101 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5161 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5221 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5281 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5341 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5401 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5461 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5521 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5581 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5641 cgaggccctt tcgtcttcac
//
