LOCUS       V006068                 7078 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006068
VERSION     V006068
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7078)
  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  JOURNAL   Unpublished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 7078)
  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 7078)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7078)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            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"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..2319
                     /gene="pol"
                     /label="DNA polymerase"
                     /note="DNA polymerase from Thermococcus gorgonarius.
                     Accession#: P56689"
     terminator      2358..2452
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             2496..3152
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     terminator      3220..3306
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(3367..3388)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3404..4483)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4482..4559)
                     /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    4751..4891
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5077..5665)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5839..6696)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6697..6801)
                     /label="AmpR promoter"
     promoter        6911..6955
                     /label="T5 promoter"
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     operon          6961..6981
                     /operon="LacO"
                     /label="LacO operon"
     protein_bind    6963..6979
                     /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             6998..7009
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             7028..7045
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             7061..7078
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
ORIGIN
        1 atgatcctcg atacagacta cataactgag gatggaaagc ccgtcatcag gatcttcaag
       61 aaggagaacg gcgagttcaa aatagactac gacagaaact ttgagccata catctacgcg
      121 ctcttgaagg acgactctgc gattgaggac gtcaagaaga taactgccga gaggcacggc
      181 actaccgtta gggttgtcag ggccgagaaa gtgaagaaga agttcctagg caggccgata
      241 gaggtctgga agctctactt cactcacccc caggacgtgc ccgcaatcag ggacaagata
      301 aaggagcatc ctgccgttgt ggacatctac gagtacgaca tccccttcgc gaagcgctac
      361 ctcatagaca aaggcttaat cccgatggag ggcgacgagg aacttaagat gctcgccttc
      421 gcgatcgcga cgctctatca cgagggcgag gagttcgccg aagggcctat cctgatgata
      481 agctacgccg acgaggaagg ggcgcgcgtt attacctgga agaatatcga ccttccctat
      541 gtcgacgtcg tttccaccga gaaggagatg ataaagcgct tcctcaaggt cgtcaaggaa
      601 aaggatcccg acgtcctcat aacctacaac ggcgacaact tcgacttcgc ctacctcaag
      661 aagcgctccg agaagctcgg agtcaagttc atcctcggaa gggaagggag cgagccgaaa
      721 atccagcgca tgggcgatcg ctttgcggtg gaggtcaagg gaaggattca cttcgacctc
      781 taccccgtca ttaggagaac gattaacctc cccacttaca cccttgaggc agtatatgaa
      841 gccatctttg gacagccgaa ggagaaggtc tacgctgagg agatagcgca ggcctgggaa
      901 acgggcgagg gattagaaag ggtggcccgc tactcgatgg aggacgcaaa ggtaacctat
      961 gaactcggaa aagagttctt ccctatggaa gcccagctct cgcgcctcgt aggccagagc
     1021 ctctgggatg tatctcgctc gagtaccgga aacctcgtcg agtggttttt gctgaggaag
     1081 gcctacgaga ggaatgaact tgcaccaaac aagccggacg agagggagct ggcaagaaga
     1141 agggagagct acgcgggtgg atacgtcaag gagcccgaaa ggggactgtg ggagaacatc
     1201 gtgtatctgg acttccgctc cctgtatcct tcgataataa tcacccataa cgtctcccct
     1261 gatacactca acagggaggg ttgtgaggag tacgacgtgg ctcctcaggt aggccataag
     1321 ttctgcaagg acttccccgg cttcatccca agcctcctcg gagacctctt ggaggagaga
     1381 cagaaggtaa agaagaagat gaaggccact atagacccaa tcgagaagaa actcctcgat
     1441 tacaggcaac gagcaatcaa aatccttgct aatagcttct acggttacta cggctatgca
     1501 aaggcccgct ggtactgcaa ggagtgcgcc gagagcgtta ccgcttgggg caggcagtac
     1561 atcgagacca cgataaggga aatagaggag aaatttggct ttaaagtcct ctacgcggac
     1621 acagatggat ttttcgcaac aatacctgga gcggacgccg aaaccgtcaa aaagaaggca
     1681 aaggagttcc tggactacat caacgccaaa ctgcccggcc tgctcgaact cgaatacgag
     1741 ggcttctaca agcgcggctt cttcgtgacg aagaagaagt acgcggttat agacgaggag
     1801 gacaagataa cgacgcgcgg gcttgaaata gttaggcgtg actggagcga gatagcgaag
     1861 gagacgcagg cgagggttct tgaggcgata ctaaagcacg gtgacgttga agaagcggta
     1921 aggattgtca aagaggttac ggagaagctg agcaagtacg aggttccacc ggagaagctg
     1981 gtcatctacg agcagataac ccgcgacctg aaggactaca aggccaccgg gccgcatgtg
     2041 gctgttgcaa aacgcctcgc cgcaaggggg ataaaaatcc ggcccggaac ggtcataagc
     2101 tacatcgtgc tcaaaggctc gggaaggatt ggggacaggg ctataccctt tgacgaattt
     2161 gacccggcaa agcacaagta cgatgcagaa tactacatcg agaaccaggt tcttccagct
     2221 gtggagagga ttctgagggc ctttggttac cgtaaagaag atttaaggta tcagaaaacg
     2281 cggcaggttg gcttgggggc gtggctaaaa cctaagacat gagtcgacct gcagccaagc
     2341 ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag aactccatct
     2401 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta
     2461 gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
     2521 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     2581 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     2641 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     2701 ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     2761 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     2821 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     2881 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     2941 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     3001 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     3061 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     3121 aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     3181 cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     3241 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     3301 acaaatccgc cgctctagat taccgtgcag tcgatgataa gctgtcaaac atgagaattg
     3361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3481 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     3541 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     3601 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     3661 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     3721 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     3781 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     3841 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     3901 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     3961 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4021 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4081 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     4141 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     4201 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     4261 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     4321 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     4381 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     4441 caccggcata ctctgcgaca tcgtataacg ttactggttt cattcaccac cctgaattga
     4501 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcg
     4561 gaatttcggg cagcgttggg tcctggccac gggtgcgcat gatctagagc tgcctcgcgc
     4621 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4681 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4741 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     4801 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     4861 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     4921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5821 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6121 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6421 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6841 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     6901 cctcgagaaa tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc
     6961 aattgtgagc ggataacaat ttcacacaga attcattaaa gaggagaaat taactatgag
     7021 aggatctcac catcaccatc accatacgga tccaagcggc ctggtgccgc gcggcagc
//
