LOCUS       V006461                 6307 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006461
VERSION     V006461
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6307)
  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  JOURNAL   Unpublished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 6307)
  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 6307)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6307)
  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            "Unpublished"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..6307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(53..241)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(1016..1037)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1053..2132)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(2133..2210)
                     /label="lacI promoter"
     promoter        2519..2537
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2538..2562
                     /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             2577..2599
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2609..2626
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             2627..3700
                     /gene="fbaA"
                     /label="Fructose-bisphosphate aldolase class 2"
                     /note="Fructose-bisphosphate aldolase class 2 from
                     Escherichia coli O157:H7. Accession#: P0AB72"
     terminator      3770..3817
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     rep_origin      3854..4309
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4405..5217)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5339..5927
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(6113..6255)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
       61 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
      121 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
      181 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
      241 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
      301 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
      361 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
      421 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
      481 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
      541 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
      601 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
      661 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
      721 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
      781 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
      841 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
      901 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
      961 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     1021 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1141 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     1201 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     1261 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     1321 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     1381 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     1441 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     1501 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     1561 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     1621 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     1681 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     1741 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     1801 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     1861 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     1921 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     1981 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     2041 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     2101 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     2161 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     2221 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     2281 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     2341 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     2401 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     2461 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
     2521 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
     2581 tttaacttta agaaggagat ataccatgca tcatcatcat catcactcta agatttttga
     2641 tttcgtaaaa cctggcgtaa tcactggtga tgacgtacag aaagttttcc aggtagcaaa
     2701 agaaaacaac ttcgcactgc cagcagtaaa ctgcgtcggt actgactcca tcaacgccgt
     2761 actggaaacc gctgctaaag ttaaagcgcc ggttatcgtt cagttctcca acggtggtgc
     2821 ttcctttatc gctggtaaag gcgtgaaatc tgacgttccg cagggtgctg ctatcctggg
     2881 cgcgatctct ggtgcgcatc acgttcacca gatggctgaa cattatggtg ttccggttat
     2941 cctgcacact gaccactgcg cgaagaaact gctgccgtgg atcgacggtc tgttggacgc
     3001 gggtgaaaaa cacttcgcag ctaccggtaa gccgctgttc tcttctcaca tgatcgacct
     3061 gtctgaagaa tctctgcaag agaacatcga aatctgctct aaatacctgg agcgcatgtc
     3121 caaaatcggc atgactctgg aaatcgaact gggttgcacc ggtggtgaag aagacggcgt
     3181 ggacaacagc cacatggacg cttctgcact gtacacccag ccggaagacg ttgattacgc
     3241 atacaccgaa ctgagcaaaa tcagcccgcg tttcaccatc gcagcgtcct tcggtaacgt
     3301 acacggtgtt tacaagccgg gtaacgtggt tctgactccg accatcctgc gtgattctca
     3361 ggaatatgtt tccaagaaac acaacctgcc gcacaacagc ctgaacttcg tattccacgg
     3421 tggttccggt tctactgctc aggaaatcaa agactccgta agctacggcg tagtaaaaat
     3481 gaacatcgat accgataccc aatgggcaac ctgggaaggc gttctgaact actacaaagc
     3541 gaacgaagct tatctgcagg gtcagctggg taacccgaaa ggcgaagatc agccgaacaa
     3601 gaaatactac gatccgcgcg tatggctgcg tgccggtcag acttcgatga tcgctcgtct
     3661 ggagaaagca ttccaggaac tgaacgcgat cgacgttctg taatgagatc cggctgctaa
     3721 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     3781 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     3841 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3901 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4201 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4261 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact
     4321 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4381 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     4441 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     4501 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     4561 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     4621 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     4681 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     4741 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     4801 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     4861 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     4921 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     4981 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     5041 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     5101 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     5161 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     5221 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     5281 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5341 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5401 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5461 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5521 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5581 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5641 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5701 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5761 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5821 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5881 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5941 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6001 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6061 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     6121 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag
     6181 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     6241 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     6301 ctgctcc
//
