LOCUS       V009608                 4731 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009608
VERSION     V009608
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4731)
  AUTHORS   Woolston BM, King JR, Reiter M, Van Hove B, Stephanopoulos G.
  TITLE     Improving formaldehyde consumption drives methanol assimilation in
            engineered E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4731)
  AUTHORS   Woolston BM, King JR, Reiter M, Van Hove B, Stephanopoulos G.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-APR-2018) Chemical Engineering, Massachusetts
            Institute of Technology, 25 Ames Street, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 4731)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4731)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (02-APR-2018) Chemical Engineering, Massachusetts Institute of
            Technology, 25 Ames Street, Cambridge, MA 02139, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(534..555)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(571..1650)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(1651..1728)
                     /label="lacI promoter"
     promoter        1774..1792
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1793..1817
                     /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             1832..1854
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1861..2868
                     /gene="glpX"
                     /label="Fructose-1,6-bisphosphatase 1 class 2"
                     /note="Fructose-1,6-bisphosphatase 1 class 2 from
                     Escherichia coli (strain K12). Accession#: P0A9C9"
     CDS             2879..2923
                     /label="S-Tag"
                     /note="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
     terminator      2975..3022
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(3245..3901)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3902..4004)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(join(4530..4731,1..343))
                     /direction=LEFT
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
ORIGIN
        1 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
       61 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
      121 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
      181 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
      241 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
      301 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
      361 tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
      421 cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag
      481 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
      541 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
      601 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
      661 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
      721 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
      781 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
      841 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
      901 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
      961 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     1021 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     1081 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     1141 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     1201 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     1261 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     1321 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     1381 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     1441 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     1501 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     1561 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     1621 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     1681 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     1741 gctctccctt atgcgactcc tgcattagga aattaatacg actcactata ggggaattgt
     1801 gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag gagatatacc
     1861 atgagacgag aacttgccat cgaattttcc cgcgtcaccg aatcagcggc gctggctggc
     1921 tacaaatggt taggacgcgg cgataaaaac accgcggacg gcgcggcggt aaacgccatg
     1981 cgtattatgc tcaaccaggt caacattgac ggcaccatcg tcattggtga aggtgaaatc
     2041 gacgaagcac cgatgctcta cattggtgaa aaagtcggta ctggtcgcgg cgacgcggta
     2101 gatattgctg ttgatccgat tgaaggcacg cgcatgacgg cgatgggcca ggctaacgcg
     2161 ctggcggtgc tggcagtagg cgataaaggc tgcttcctca atgcgccgga tatgtatatg
     2221 gagaagctga ttgtcgggcc gggagccaaa ggcaccattg atctgaacct gccgctggcg
     2281 gataacctgc gcaatgtagc ggcggcgctc ggcaaaccgt tgagcgaact gacggtaacg
     2341 attctggcta aaccacgcca cgatgccgtt atcgctgaaa tgcagcaact cggcgtacgc
     2401 gtatttgcta ttccggacgg cgacgttgcg gcctcaattc tcacctgtat gccagacagc
     2461 gaagttgacg tgctgtacgg tattggtggc gcgccggaag gcgtagtttc tgcggcggtg
     2521 atccgcgcat tagatggcga catgaacggt cgtctgctgg cgcgtcatga cgtcaaaggc
     2581 gacaacgaag agaatcgtcg cattggcgag caggagctgg cacgctgcaa agcgatgggc
     2641 atcgaagccg gtaaagtatt gcgcctgggc gatatggcgc gcagcgataa cgtcatcttc
     2701 tctgccaccg gtattaccaa aggcgatctg ctggaaggca ttagccgcaa aggcaatatc
     2761 gcgactaccg aaacgctgct gatccgcggc aagtcacgca ccattcgccg cattcagtcc
     2821 atccactatc tggatcgcaa agacccggaa atgcaggtgc acatcctctg agtctggtaa
     2881 agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta
     2941 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     3001 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
     3061 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     3121 ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta
     3181 ttatcactta ttcaggcgta gcaccaggcg tttaagggca ccaataactg ccttaaaaaa
     3241 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     3301 catggaagcc atcacagacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     3361 cgccttgcgt ataatatttg cccatagtga aaacgggggc gaagaagttg tccatattgg
     3421 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     3481 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     3541 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     3601 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     3661 caccgtcttt cattgccata cggaactccg gatgagcatt catcaggcgg gcaagaatgt
     3721 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     3781 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     3841 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     3901 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     3961 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc
     4021 aaaagttggc ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga
     4081 agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat gctgccaact
     4141 tactgattta gtgtatgatg gtgtttttga ggtgctccag tggcttctgt ttctatcagc
     4201 tgtccctcct gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac cgccggacat
     4261 cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
     4321 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
     4381 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     4441 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     4501 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     4561 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     4621 gcgtttcccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     4681 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag t
//
