LOCUS       V015767                 6169 bp    DNA     circular SYN 12-DEC-2019
DEFINITION  Exported.
ACCESSION   V015767
VERSION     V015767
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6169)
  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   2  (bases 1 to 6169)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (10-MAY-2019) Biocatalysis "
FEATURES             Location/Qualifiers
     source          1..6169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..88
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             120..142
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             161..178
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             188..205
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             209..1699
                     /gene="glpK"
                     /label="Glycerol kinase"
                     /note="Glycerol kinase from Thermococcus kodakarensis
                     (strain ATCC BAA-918 / JCM 12380 / KOD1). Accession#:
                     O93623"
     CDS             1757..2884
                     /gene="ackA"
                     /label="Acetate kinase"
                     /note="Acetate kinase from Mycolicibacterium smegmatis
                     (strain ATCC 700084 / mc(2)155). Accession#: A0QQK1"
     terminator      3881..3928
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        complement(4294..4322)
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        4435..4539
                     /label="AmpR promoter"
     CDS             4540..5397
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5571..6159
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg gatctcgatc
       61 ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct agaaataatt
      121 ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag
      181 cagcggcctg gtgccgcgcg gcagccatat ggaaaaattt gtgctgagcc tggatgaagg
      241 tacaaccagc gcacgtgcaa ttatctttga tcgtgaaagc aacattcatg gcattggcca
      301 gtatgaattt ccgcagcatt atccgcgtcc gggttgggtt gaacataatc cggaagaaat
      361 ttgggatgca cagctgcgtg ccattaaaga tgcaattcag agcgcacgta ttgaaccgaa
      421 tcagattgca gcaattggtg ttaccaatca gcgtgaaacc accctggttt gggataaaga
      481 tggtaaaccg ctgtataatg caattgtttg gcagtgtcgt cgtaccgcag aaatggttga
      541 agaaattaaa cgtgaatacg gcaccatgat caaagaaaaa accggtctgg ttccggatgc
      601 atattttagc gcaagcaaac tgaaatggct gctggataat gttccgggtc tgcgtgaaaa
      661 agccgaaaaa ggtgaagtta tgtttggcac cgttgatacc tttctgattt atcgtctgac
      721 cggtgaacat gtgaccgatt atagcaatgc aagccgtacc atgctgttta acatcaaaaa
      781 actggattgg gatgatgagc tgctggaact gtttgatatt ccggaaagcg ttctgccgga
      841 agttcgtgaa agtagcgaag tttatggcta taccaaaaaa gaactgctgg gtgcagaaat
      901 tccggttagc ggtgatgccg gtgatcagca ggcagcactg tttggtcagg cagcatttga
      961 agcaggtatg gttaaagcaa cctatggcac cggtagcttt attctggtta ataccgataa
     1021 aatggtcctg tatagcgata atctgctgac caccattgca tggggtctga atggtcgtgt
     1081 tagctatgca ctggaaggta gcatttttgt taccggtgca gcagttcagt ggctgcgtga
     1141 tggcattaaa atcattaaac atgccagcga aaccgaagaa ctggcaacca aactggaaag
     1201 caatgaaggt gtttattttg tgcctgcatt tgttggtctg ggtgcaccgt attgggatca
     1261 gtttgcacgt ggtattatca ttggtattac ccgtggtaca ggtcgtgaac atctggcacg
     1321 tgcgaccctg gaagcaattg catatctgac ccgtgatgtt gttgatgaga tggaaaaact
     1381 ggtgcagatt aaagaactgc gtgttgatgg tggtgcaacc gcaaatgatt ttctgatgca
     1441 gtttcaggcc gatattctga atcgtaaagt tattcgtccg gtggtgaaag aaaccaccgc
     1501 actgggtgcc gcatatctgg caggtctggc agttgattat tgggcagata cccgtgaaat
     1561 tgcagaactg tggaaagcag aacgtatttt tgagccgaaa atggatgaaa aaacccgtga
     1621 acgtctgtac aaaggttgga aagaagcagt taaacgcgca atgggttggg caaaagttgt
     1681 tgatagcgca aaaagcaatg gtagcagcgg tagctcaggt agcagctgcg gtagcagtgg
     1741 ttcaagcggt agtagcatga ccgttctggt tgttaatagc ggttcaagca gcctgaaata
     1801 tgcagttgtg cgtccggcaa gcggtgaatt tctggcagat ggtattattg aagagattgg
     1861 tagcggtgca gttccggatc atgatgcagc cctgcgtgca gcctttgatg aactggcagc
     1921 agcaggtctg catctggaag atctggatct gaaagcagtt ggtcatcgta tggttcatgg
     1981 tggtaaaacc ttctataaac cgagcgttgt tgacgatgaa ctgattgcaa aagcacgtga
     2041 actgagtccg ctggcaccgc tgcataatcc gcctgcaatt aaaggtattg aagttgcacg
     2101 taaactgctg ccggatctgc ctcatattgc agtttttgat accgcctttt ttcatgatct
     2161 gcctgcaccg gcaagcacct atgccattga tcgcgaactg gccgaaacct ggcatattaa
     2221 acgttatggt tttcatggca ccagccatga atatgttagc cagcaagcag caatttttct
     2281 ggatcgtccg ctggaaagcc tgaaccagat tgttctgcac ctgggtaatg gtgcaagcgc
     2341 cagcgcagtt gccggtggta aagcggttga taccagcatg ggtctgaccc cgatggaagg
     2401 tctggttatg ggcacccgta gtggtgatat tgatccgggt gttattatgt atctgtggcg
     2461 caccgcaggt atgagcgttg atgatattga aagtatgctg aatcgtcgta gcggtgttct
     2521 gggcctgggt ggtgcgagcg attttcgcaa actgcgcgaa ctgatcgaaa gtggtgatga
     2581 acatgcaaaa ctggcctatg atgtgtatat tcatcgtctg cgcaaatata tcggtgcata
     2641 tatggccgtt ctgggtcgta ccgatgttat tagctttacc gcaggcgttg gtgaaaatgt
     2701 tccgcctgtt cgtcgtgatg ccctggcagg actgggtggt ctgggcattg aaattgatga
     2761 tgcactgaat agcgccaaaa gtgatgaacc gcgtctgatt agcacaccgg atagccgtgt
     2821 tacagttctg gtggttccga ccaatgagga actggccatt gcccgtgcat gtgttggtgt
     2881 tgttggtacc ggtagtagtg gtagtagcgg cagcagcggc tcaagcatgc cgctggaccc
     2941 ggttattcag caggttctgg atcagctgaa ccgtatgcca gcaccggatt ataaacatct
     3001 gagcgcacag cagtttcgta gccagcagag cctgtttcct ccggttaaaa aagagccggt
     3061 tgcagaagtt cgcgaatttg atatggatct gccaggtcgt accctgaaag ttcgtatgta
     3121 tcgtccggaa ggtgttgaac cgccttatcc ggcactggtt tattatcatg gtggcggttg
     3181 ggtggttggt gatctggaaa cccatgatcc tgtttgtcgt gttctggcaa aagatggtcg
     3241 tgcagttgtt tttagcgttg attatcgcct ggcaccggaa cataaatttc cggcagccgt
     3301 tgaagatgcg tatgatgcgc tgcagtggat tgccgaacgt gcagcagatt ttcatctgga
     3361 ccctgcccgt attgccgttg gtggtgatag tgcgggtggt aatctggcag cggttaccag
     3421 cattctggcc aaagaacgtg gtggtcctgc actggcattt cagctgctga tctatccgag
     3481 caccggttat gatccggcac atccgccagc cagcattgaa gaaaatgcag aaggctacct
     3541 gctgacaggt ggtatgatgc tgtggtttcg tgatcagtat ctgaatagcc tggaagaact
     3601 gacccatccg tggtttagtc cggttctgta tccggatctg agcggtctgc ctccggcata
     3661 tattgccacc gcacagtatg atcctctgcg tgacgttggt aaactgtatg cagaagccct
     3721 gaataaagcc ggtgttaaag tggaaatcga aaactttgaa gatctgatcc atggctttgc
     3781 ccagttttat agcctgagtc cgggtgcaac caaagcactg gttcgtattg cagaaaaact
     3841 gcgggatgca ctggcctaat aaggatccgc tgagcaataa ctagcataac cccttggggc
     3901 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca
     3961 ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca
     4021 ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt
     4081 gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga
     4141 cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca
     4201 gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac
     4261 tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc
     4321 aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4381 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4441 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4501 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4561 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4621 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4681 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4741 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     4801 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4861 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4921 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4981 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5041 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     5101 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5161 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5221 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5281 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5341 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5401 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5461 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5521 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5701 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5941 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagca
//
