LOCUS       40924_21270        7081 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pGDR11-KOD, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7081)
  AUTHORS   Dunn MR, Otto CE, Fenton KE, Chaput JC.
  TITLE     Transference as a Mechanism for Expanding and Improving Polymerase 
            Function
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7081)
  AUTHORS   Dunn MR, Otto CE, Fenton KE, Chaput JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-2015) The Biodesign Institute, Arizona State 
            University, 727 E Tyler Street, Tempe, AZ 85287, USA
REFERENCE   3  (bases 1 to 7081)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7081)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-JAN-2015) The Biodesign Institute, Arizona State University, 727
            E Tyler Street, Tempe, AZ 85287, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..2325
                     /codon_start=1
                     /gene="pol"
                     /product="exonuclease-deficient DNA polymerase"
                     /label=pol
                     /note="from Thermococcus kodakarensis"
                     /protein_id="ALL53339.1"
                     /translation="MILDTDYITEDGKPVIRIFKKENGEFKIEYDRTFEPYFYALLKDD
                     SAIEEVKKITAERHGTVVTVKRVEKVQKKFLGRPVEVWKLYFTHPQDVPAIRDKIREHP
                     AVIDIYEYDIPFAKRYLIDKGLVPMEGDEELKMLAFAIATLYHEGEEFAEGPILMISYA
                     DEEGARVITWKNVDLPYVDVVSTEREMIKRFLRVVKEKDPDVLITYNGDNFDFAYLKKR
                     CEKLGINFALGRDGSEPKIQRMGDRFAVEVKGRIHFDLYPVIRRTINLPTYTLEAVYEA
                     VFGQPKEKVYAEEITTAWETGENLERVARYSMEDAKVTYELGKEFLPMEAQLSRLIGQS
                     LWDVSRSSTGNLVEWFLLRKAYERNELAPNKPDEKELARRRQSYEGGYVKEPERGLWEN
                     IVYLDFRSLYPSIIITHNVSPDTLNREGCKEYDVAPQVGHRFCKDFPGFIPSLLGDLLE
                     ERQKIKKKMKATIDPIERKLLDYRQRAIKILANSYYGYYGYARARWYCKECAESVTAWG
                     REYITMTIKEIEEKYGFKVIYSDTDGFFATIPGADAETVKKKAMEFLKYINAKLPGALE
                     LEYEGFYKRGFFVTKKKYAVIDEEGKITTRGLEIVRRDWSEIAKETQARVLEALLKDGD
                     VEKAVRIVKEVTEKLSKYEVPPEKLVIHEQITRDLKDYKATGPHVAVAKRLAARGVKIR
                     PGTVISYIVLKGSGRIGDRAIPFDEFDPTKHKYDAEYYIENQVLPAVERILRAFGYRKE
                     DLRYQKTRQVGLSAWLKPKGT"
     gene            1..2325
                     /gene="pol"
                     /label=pol
     terminator      2361..2455
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2499..3155
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      3223..3309
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(3370..3391)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3407..4486)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4485..4562)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    4754..4894
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5080..5668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5842..6699)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6700..6804)
                     /label=AmpR promoter
     promoter        6914..6958
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     operon          6964..6984
                     /operon="LacO"
                     /label=LacO operon
     protein_bind    6966..6982
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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             7001..7012
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7031..7048
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             7064..7081
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
ORIGIN
        1 atgatcctcg acactgacta cataaccgag gatggaaagc ctgtcataag aattttcaag
       61 aaggaaaacg gcgagtttaa gattgagtac gaccggactt ttgaacccta cttctacgcc
      121 ctcctgaagg acgattctgc cattgaggaa gtcaagaaga taaccgccga gaggcacggg
      181 acggttgtaa cggttaagcg ggttgaaaag gttcagaaga agttcctcgg gagaccagtt
      241 gaggtctgga aactctactt tactcatccg caggacgtcc cagcgataag ggacaagata
      301 cgagagcatc cagcagttat tgacatctac gagtacgaca tacccttcgc caagcgctac
      361 ctcatagaca agggattagt gccaatggaa ggcgacgagg agctgaaaat gctcgccttc
      421 gcgattgcga ctctctacca tgagggcgag gagttcgccg aggggccaat ccttatgata
      481 agctacgccg acgaggaagg ggccagggtg ataacttgga agaacgtgga tctcccctac
      541 gttgacgtcg tctcgacgga gagggagatg ataaagcgct tcctccgtgt tgtgaaggag
      601 aaagacccgg acgttctcat aacctacaac ggcgacaact tcgacttcgc ctatctgaaa
      661 aagcgctgtg aaaagctcgg aataaacttc gccctcggaa gggatggaag cgagccgaag
      721 attcagagga tgggcgacag gtttgccgtc gaagtgaagg gacggataca cttcgatctc
      781 tatcctgtga taagacggac gataaacctg cccacataca cgcttgaggc cgtttatgaa
      841 gccgtcttcg gtcagccgaa ggagaaggtt tacgctgagg aaataaccac agcctgggaa
      901 accggcgaga accttgagag agtcgcccgc tactcgatgg aagatgcgaa ggtcacatac
      961 gagcttggga aggagttcct tccgatggag gcccagcttt ctcgcttaat cggccagtcc
     1021 ctctgggacg tctcccgctc cagcactggc aacctcgttg agtggttcct cctcaggaag
     1081 gcctatgaga ggaatgagct ggccccgaac aagcccgatg aaaaggagct ggccagaaga
     1141 cggcagagct atgaaggagg ctatgtaaaa gagcccgaga gagggttgtg ggagaacata
     1201 gtgtacctag attttagatc cctgtacccc tcaatcatca tcacccacaa cgtctcgccg
     1261 gatacgctca acagagaagg atgcaaggaa tatgacgttg ccccacaggt cggccaccgc
     1321 ttctgcaagg acttcccagg atttatcccg agcctgcttg gagacctcct agaggagagg
     1381 cagaagataa agaagaagat gaaggccacg attgacccga tcgagaggaa gctcctcgat
     1441 tacaggcaga gggccatcaa gatcctggca aacagctact acggttacta cggctatgca
     1501 agggcgcgct ggtactgcaa ggagtgtgca gagagcgtaa cggcctgggg aagggagtac
     1561 ataacgatga ccatcaagga gatagaggaa aagtacggct ttaaggtaat ctacagcgac
     1621 accgacggat tttttgccac aatacctgga gccgatgctg aaaccgtcaa aaagaaggct
     1681 atggagttcc tcaagtatat caacgccaaa cttccgggcg cgcttgagct cgagtacgag
     1741 ggcttctaca aacgcggctt cttcgtcacg aagaagaagt atgcggtgat agacgaggaa
     1801 ggcaagataa caacgcgcgg acttgagatt gtgaggcgtg actggagcga gatagcgaaa
     1861 gagacgcagg cgagggttct tgaagctttg ctaaaggacg gtgacgtcga gaaggccgtg
     1921 aggatagtca aagaagttac cgaaaagctg agcaagtacg aggttccgcc ggagaagctg
     1981 gtgatccacg agcagataac gagggattta aaggactaca aggcaaccgg tccccacgtt
     2041 gccgttgcca agaggttggc cgcgagagga gtcaaaatac gccctggaac ggtgataagc
     2101 tacatcgtgc tcaagggctc tgggaggata ggcgacaggg cgataccgtt cgacgagttc
     2161 gacccgacga agcacaagta cgacgccgag tactacattg agaaccaggt tctcccagcc
     2221 gttgagagaa ttctgagagc cttcggttac cgcaaggaag acctgcgcta ccagaagacg
     2281 agacaggttg gtttgagtgc ttggctgaag ccgaagggaa cttgagtcga cctgcagcca
     2341 agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     2401 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
     2461 ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     2521 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     2581 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     2641 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     2701 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     2761 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     2821 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     2881 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     2941 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     3001 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     3061 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     3121 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     3181 gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct
     3241 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     3301 aggacaaatc cgccgctcta gattaccgtg cagtcgatga taagctgtca aacatgagaa
     3361 ttgtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3421 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3481 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     3541 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     3601 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     3661 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     3721 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     3781 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     3841 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     3901 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     3961 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     4021 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     4081 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     4141 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     4201 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     4261 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     4321 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     4381 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     4441 agacaccggc atactctgcg acatcgtata acgttactgg tttcattcac caccctgaat
     4501 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg
     4561 tcggaatttc gggcagcgtt gggtcctggc cacgggtgcg catgatctag agctgcctcg
     4621 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4681 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4741 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     4801 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     4861 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     4921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5521 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6121 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6421 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6841 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6901 tcacctcgag aaatcataaa aaatttattt gctttgtgag cggataacaa ttataataga
     6961 ttcaattgtg agcggataac aatttcacac agaattcatt aaagaggaga aattaactat
     7021 gagaggatct caccatcacc atcaccatac ggatccaagc ggcctggtgc cgcgcggcag
     7081 c
//
