LOCUS       V015888                 5197 bp    DNA     circular SYN 12-DEC-2019
DEFINITION  Exported.
ACCESSION   V015888
VERSION     V015888
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5197)
  AUTHORS   Hartley CJ, Williams CC, Scoble JA, Churches QI, North A, French NG,
            Coia G, Nebl T, Warden AC, Simpson G, Frazer AR, Nixon Jensen C,
            Turner NJ, Scott C.
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5197)
  AUTHORS   Hartley CJ, Williams CC, Scoble JA, Churches QI, North A, French NG,
            Nebl T, Coia G, Warden AC, Simpson G, Frazer AR, Nixon Jensen C,
            Turner NJ, Scott C.
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5197)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (10-MAY-2019) Biocatalysis "
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (10-MAY-2019) Biocatalysis "
FEATURES             Location/Qualifiers
     source          1..5197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2..1492
                     /gene="glpK"
                     /label="Glycerol kinase"
                     /note="Glycerol kinase from Thermococcus kodakarensis
                     (strain ATCC BAA-918 / JCM 12380 / KOD1). Accession#:
                     O93623"
     CDS             1550..2677
                     /gene="ackA"
                     /label="Acetate kinase"
                     /note="Acetate kinase from Mycolicibacterium smegmatis
                     (strain ATCC 700084 / mc(2)155). Accession#: A0QQK1"
     terminator      2702..2749
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        complement(3115..3143)
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        3256..3360
                     /label="AmpR promoter"
     CDS             3361..4218
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4392..4980
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        5060..5078
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             5110..5132
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5151..5168
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             5178..5195
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
ORIGIN
        1 tatggaaaaa tttgtgctga gcctggatga aggtacaacc agcgcacgtg caattatctt
       61 tgatcgtgaa agcaacattc atggcattgg ccagtatgaa tttccgcagc attatccgcg
      121 tccgggttgg gttgaacata atccggaaga aatttgggat gcacagctgc gtgccattaa
      181 agatgcaatt cagagcgcac gtattgaacc gaatcagatt gcagcaattg gtgttaccaa
      241 tcagcgtgaa accaccctgg tttgggataa agatggtaaa ccgctgtata atgcaattgt
      301 ttggcagtgt cgtcgtaccg cagaaatggt tgaagaaatt aaacgtgaat acggcaccat
      361 gatcaaagaa aaaaccggtc tggttccgga tgcatatttt agcgcaagca aactgaaatg
      421 gctgctggat aatgttccgg gtctgcgtga aaaagccgaa aaaggtgaag ttatgtttgg
      481 caccgttgat acctttctga tttatcgtct gaccggtgaa catgtgaccg attatagcaa
      541 tgcaagccgt accatgctgt ttaacatcaa aaaactggat tgggatgatg agctgctgga
      601 actgtttgat attccggaaa gcgttctgcc ggaagttcgt gaaagtagcg aagtttatgg
      661 ctataccaaa aaagaactgc tgggtgcaga aattccggtt agcggtgatg ccggtgatca
      721 gcaggcagca ctgtttggtc aggcagcatt tgaagcaggt atggttaaag caacctatgg
      781 caccggtagc tttattctgg ttaataccga taaaatggtc ctgtatagcg ataatctgct
      841 gaccaccatt gcatggggtc tgaatggtcg tgttagctat gcactggaag gtagcatttt
      901 tgttaccggt gcagcagttc agtggctgcg tgatggcatt aaaatcatta aacatgccag
      961 cgaaaccgaa gaactggcaa ccaaactgga aagcaatgaa ggtgtttatt ttgtgcctgc
     1021 atttgttggt ctgggtgcac cgtattggga tcagtttgca cgtggtatta tcattggtat
     1081 tacccgtggt acaggtcgtg aacatctggc acgtgcgacc ctggaagcaa ttgcatatct
     1141 gacccgtgat gttgttgatg agatggaaaa actggtgcag attaaagaac tgcgtgttga
     1201 tggtggtgca accgcaaatg attttctgat gcagtttcag gccgatattc tgaatcgtaa
     1261 agttattcgt ccggtggtga aagaaaccac cgcactgggt gccgcatatc tggcaggtct
     1321 ggcagttgat tattgggcag atacccgtga aattgcagaa ctgtggaaag cagaacgtat
     1381 ttttgagccg aaaatggatg aaaaaacccg tgaacgtctg tacaaaggtt ggaaagaagc
     1441 agttaaacgc gcaatgggtt gggcaaaagt tgttgatagc gcaaaaagca atggtagcag
     1501 cggtagctca ggtagcagct gcggtagcag tggttcaagc ggtagtagca tgaccgttct
     1561 ggttgttaat agcggttcaa gcagcctgaa atatgcagtt gtgcgtccgg caagcggtga
     1621 atttctggca gatggtatta ttgaagagat tggtagcggt gcagttccgg atcatgatgc
     1681 agccctgcgt gcagcctttg atgaactggc agcagcaggt ctgcatctgg aagatctgga
     1741 tctgaaagca gttggtcatc gtatggttca tggtggtaaa accttctata aaccgagcgt
     1801 tgttgacgat gaactgattg caaaagcacg tgaactgagt ccgctggcac cgctgcataa
     1861 tccgcctgca attaaaggta ttgaagttgc acgtaaactg ctgccggatc tgcctcatat
     1921 tgcagttttt gataccgcct tttttcatga tctgcctgca ccggcaagca cctatgccat
     1981 tgatcgcgaa ctggccgaaa cctggcatat taaacgttat ggttttcatg gcaccagcca
     2041 tgaatatgtt agccagcaag cagcaatttt tctggatcgt ccgctggaaa gcctgaacca
     2101 gattgttctg cacctgggta atggtgcaag cgccagcgca gttgccggtg gtaaagcggt
     2161 tgataccagc atgggtctga ccccgatgga aggtctggtt atgggcaccc gtagtggtga
     2221 tattgatccg ggtgttatta tgtatctgtg gcgcaccgca ggtatgagcg ttgatgatat
     2281 tgaaagcatg ctgaatcgtc gtagcggtgt tctgggcctg ggtggtgcga gcgattttcg
     2341 caaactgcgc gaactgatcg aaagtggtga tgaacatgca aaactggcct atgatgtgta
     2401 tattcatcgt ctgcgcaaat atatcggtgc atatatggcc gttctgggtc gtaccgatgt
     2461 tattagcttt accgcaggcg ttggtgaaaa tgttccgcct gttcgtcgtg atgccctggc
     2521 aggactgggt ggtctgggca ttgaaattga tgatgcactg aatagcgcca aaagtgatga
     2581 accgcgtctg attagcacac cggatagccg tgttacagtt ctggtggttc cgaccaatga
     2641 ggaactggcc attgcccgtg catgtgttgg tgttgtttaa tgaggatccg ctgagcaata
     2701 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     2761 aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg
     2821 ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga
     2881 gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac
     2941 tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca
     3001 agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag
     3061 atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa
     3121 gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata
     3181 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3241 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3301 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3361 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3421 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3481 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3541 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3601 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3661 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3721 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3781 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3841 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3901 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3961 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4021 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4081 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4141 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4201 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4261 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4321 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4381 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4441 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4501 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     4561 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4621 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4681 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4741 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4801 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4861 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4921 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4981 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5041 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
     5101 agaaataatt ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc
     5161 atcatcacag cagcggcctg gtgccgcgcg gcagcca
//
