LOCUS       V006460                 6238 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006460
VERSION     V006460
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6238)
  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  JOURNAL   Unpublished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 6238)
  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 6238)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6238)
  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            "Unpublished"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..6238
                     /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..3631
                     /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"
     terminator      3701..3748
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     rep_origin      3785..4240
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4336..5148)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5270..5858
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(6044..6186)
                     /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 catcacagac gagaacttgc
     2641 catcgaattt tcccgcgtca ccgaatcagc ggcgctggct ggctacaaat ggttaggacg
     2701 cggcgataaa aacaccgcgg acggcgcggc ggtaaacgcc atgcgtatta tgctcaacca
     2761 ggtcaacatt gacggcacca tcgtcattgg tgaaggtgaa atcgacgaag caccgatgct
     2821 ctacattggt gaaaaagtcg gtactggtcg cggcgacgcg gtagatattg ctgttgatcc
     2881 gattgaaggc acgcgcatga cggcgatggg ccaggctaac gcgctggcgg tgctggcagt
     2941 aggcgataaa ggctgcttcc tcaatgcgcc ggatatgtat atggagaagc tgattgtcgg
     3001 gccgggagcc aaaggcacca ttgatctgaa cctgccgctg gcggataacc tgcgcaatgt
     3061 agcggcggcg ctcggcaaac cgttgagcga actgacggta acgattctgg ctaaaccacg
     3121 ccacgatgcc gttatcgctg aaatgcagca actcggcgta cgcgtatttg ctattccgga
     3181 cggcgacgtt gcggcctcaa ttctcacctg tatgccagac agcgaagttg acgtgctgta
     3241 cggtattggt ggcgcgccgg aaggcgtagt ttctgcggcg gtgatccgcg cattagatgg
     3301 cgacatgaac ggtcgtctgc tggcgcgtca tgacgtcaaa ggcgacaacg aagagaatcg
     3361 tcgcattggc gagcaggagc tggcacgctg caaagcgatg ggcatcgaag ccggtaaagt
     3421 attgcgcctg ggcgatatgg cgcgcagcga taacgtcatc ttctctgcca ccggtattac
     3481 caaaggcgat ctgctggaag gcattagccg caaaggcaat atcgcgacta ccgaaacgct
     3541 gctgatccgc ggcaagtcac gcaccattcg ccgcattcag tccatccact atctggatcg
     3601 caaagacccg gaaatgcagg tgcacatcct ctgatgagat ccggctgcta acaaagcccg
     3661 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     3721 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     3781 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3841 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4081 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4201 tttaacgcga attttaacaa aatattaacg tttacaattt caggtggcac ttttcgggga
     4261 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4321 atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     4381 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     4441 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     4501 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     4561 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     4621 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     4681 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     4741 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     4801 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     4861 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     4921 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     4981 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     5041 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     5101 atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     5161 attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt
     5221 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5281 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5341 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5401 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5461 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5521 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5581 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5641 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5701 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5761 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5821 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5881 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5941 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6001 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     6061 tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg
     6121 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     6181 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcc
//
