LOCUS       V006281                 8509 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006281
VERSION     V006281
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8509)
  AUTHORS   Farzadfard F, Lu TK.
  TITLE     Synthetic biology. Genomically encoded analog memory with precise in
            vivo DNA writing in living cell populations
  JOURNAL   Science 346 (6211), 1256272 (2014)
   PUBMED   25395541
REFERENCE   2  (bases 1 to 8509)
  AUTHORS   Farzadfard F, Lu T.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2014) Biological Engineering, MIT, 77
            Massachusetts Avenue, NE47, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 8509)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8509)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science";
            date: "2014"; volume: "346"; issue: "6211"; pages: "1256272"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (04-DEC-2014) Biological Engineering, MIT, 77 Massachusetts Avenue,
            NE47, Cambridge, MA 02139, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(42..184)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(289..477)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(1281..1895)
                     /gene="fixJ"
                     /label="Transcriptional regulatory protein FixJ"
                     /note="Transcriptional regulatory protein FixJ from
                     Bradyrhizobium diazoefficiens (strain JCM 10833 / BCRC
                     13528 / IAM 13628 / NBRC 14792 / USDA 110). Accession#:
                     P23221"
     CDS             complement(1892..3034)
                     /codon_start=1
                     /product="YF1"
                     /label="YF1"
                     /protein_id="AIZ09121.1"
                     /translation="MASFQSFGIPGQLEVIKKALDHVRVGVVITDPALEDNPIVYVNQG
                     FVQMTGYETEEILGKNCRFLQGKHTDPAEVDNIRTALQNKEPVTVQIQNYKKDGTMFWN
                     ELNIDPMEIEDKTYFVGIQNDITEHQQTQARLQELQSELVHVSRLSAMGEMASALAHEL
                     NQPLAAISNYMKGSRRLLAGSSDPNTPKVESALDRAAEQALRAGQIIRRLRDFVARGES
                     EKRVESLSKLIEEAGALGLAGAREQNVQLRFSLDPGADLVLADRVQIQQVLVNLFRNAL
                     EAMAQSQRRELVVTNTPAADDMIEVEVSDTGSGFQDDVIPNLFQTFFTTKDTGMGVGLS
                     ISRSIIEAHGGRMWAESNASGGATFRFTLPAADEMIGGLA"
     promoter        complement(3035..3112)
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     regulatory      3407..3653
                     /label="FixK2 promoter"
                     /note="FixK2 promoter"
                     /regulatory_class="promoter"
     RBS             3660..3671
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             3678..4388
                     /label="lambda repressor"
                     /note="phage lambda repressor"
     CDS             4389..4421
                     /label="ssrA tag (LVA)"
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness
                     et al., 2006)"
     terminator      4469..4540
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4556..4583
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     regulatory      4598..4646
                     /label="lambda pR promoter"
                     /note="lambda pR promoter"
                     /regulatory_class="promoter"
     misc_feature    4655..4736
                     /note="primer for the Ec86 RT; msr"
     misc_feature    complement(4726..4856)
                     /note="template for Ec86 RT; msd(kanR_ON)"
     CDS             4876..5835
                     /gene="ret"
                     /label="Retron Ec86 reverse transcriptase"
                     /note="Retron Ec86 reverse transcriptase from Escherichia
                     coli. Accession#: P23070"
     CDS             5875..6657
                     /label="Beta"
                     /note="single-stranded DNA binding recombinase in the
                     lambda Red system"
     terminator      6678..6764
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(6928..7516)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     terminator      complement(7604..7698)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(7722..8378)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(8379..8481)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
ORIGIN
        1 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
       61 cttacgcatc tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct
      121 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
      181 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      241 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      301 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
      361 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
      421 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
      481 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
      541 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
      601 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
      661 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
      721 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
      781 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
      841 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
      901 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
      961 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     1021 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     1081 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     1141 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     1201 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     1261 ctaacttaca tcaattgtca atcgttgagc atgccggcgc gcatcgcgag gcgaaccagc
     1321 tccgaaaggc tgttggcctg catcttggtc atgacgttgg cccgatacac ctcgatggtg
     1381 cgcgggctga tgtcgtactc gcgggcgatc agcttgttgg aaaggccggc gatcagccct
     1441 tccatgacct ggcgctccct ggggctcaac gaggcgacgc gggcggcgat atcctgcgcg
     1501 acggcctcgc tcttggcggc cggctcggcc tggcggatcg ccgattcgat catggcggtg
     1561 aggcggtcgt cctcgaaagg cttttccaga aagtccaccg cccctaactt catcgcctcg
     1621 accgcgagcg gcacgtcgcc gtgaccggtc atgatgagga tcggaaaggg gctttgctgc
     1681 gccttcatcc gcttcaacag ctcgatgccg tcaaggcccg gcatgcgcac gtcggagacg
     1741 acgcagccga aggagagacc cggcagggcg tcgagaaagg cttgcgcgtc gtcaaacagc
     1801 gtgacgccga agccggcaga atccagcagg aaattcagcg aatcccgcat cgccgcgtcg
     1861 tcgtcgatga cgtagatatg tcccttggtc gtcatgctag acctcctatc atctcgtcgg
     1921 ctgccgggag ggtgaagcgg aaggtcgccc cgcccgatgc gttgctctcg gcccacatgc
     1981 gcccgccgtg agcttcgatg atcgagcggc tgatggacag tcccacgccc atgccggtgt
     2041 ccttggtggt gaagaaagtc tgaaacaggt tcggaatgac gtcgtcctgg aaaccgctgc
     2101 cggtgtcgga cacttcgacc tcgatcatgt cgtcggcggc gggggtgttg gtgacgacga
     2161 gctcgcgtcg ctgcgactga gccatcgctt ccagcgcgtt gcggaacagg ttgaccagga
     2221 cctgctggat ctgcacccgg tcggcgagaa cgagatcggc gcccggatcg agactgaagc
     2281 ggagctgcac gttctgctcg cgcgcgccgg caagcccgag cgcgccggcc tcctcgatca
     2341 gcttggagag actctcgacc cgcttctccg attcgccgcg ggcaacgaag tcgcgcaggc
     2401 gccggatgat ctggccggcg cgcagcgcct gctcggcggc gcggtccagg gcgctttcga
     2461 ccttcggtgt gttcggatca ctgctgccgg caagcagccg ccgcgagccc ttcatgtagt
     2521 tgctgatcgc cgccagcggc tggttgagct cgtgcgcgag cgcggacgcc atttcgccca
     2581 tggcgctcag cctggagacg tggacgagct cggattgcag ttcctggagg cgcgcctggg
     2641 tctgctggtg ctcggtgata tcattctgaa taccgacaaa atacgtttta tcctctattt
     2701 ccattggatc aatatttaat tcattccaga acatcgttcc gtcttttttg tagttttgga
     2761 tctgaacggt gaccggttct ttattttgta aagcggttct gatgttgtcc acttctgcag
     2821 gatctgtgtg tttcccctgt aagaagcgac agttctttcc taaaatttcc tcggtctcgt
     2881 agccggtcat ttgaacaaag ccttgattta cgtagacaat aggattatct tcaagtgcgg
     2941 gatctgtaat taccacaccg actcgcacgt gatcaagtgc ttttttgatg acttccagct
     3001 gtcctggtat cccaaatgat tgaaaactag ccatattcac caccctcaat tgactctctt
     3061 ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tccgggatct
     3121 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     3181 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     3241 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     3301 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     3361 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctacgc ccgtgatcct
     3421 gatcaccggc tatccggacg aaaacatctc gacccgggcc gccgaggccg gcgtaaaaga
     3481 cgtggttttg aagccgcttc tcgacgaaaa cctgctcaag cgtatccgcc gcgccatcca
     3541 ggaccggcct cgggcatgac ctacggggtt ctacgtaagg cacccccctt aagatatcgc
     3601 tcgaaatttt cgaacctccc gataccgcgt accaatgcgt catcacaacg gagtctagaa
     3661 aagaggagaa atactagatg agcacaaaaa agaaaccatt aacacaagag cagcttgagg
     3721 acgcacgtcg ccttaaagca atttatgaaa aaaagaaaaa tgaacttggc ttatcccagg
     3781 aatctgtcgc agacaagatg gggatggggc agtcaggcgt tggtgcttta tttaatggca
     3841 tcaatgcatt aaatgcttat aacgccgcat tgcttgcaaa aattctcaaa gttagcgttg
     3901 aagaatttag cccttcaatc gccagagaaa tctacgagat gtatgaagcg gttagtatgc
     3961 agccgtcact tagaagtgag tatgagtacc ctgttttttc tcatgttcag gcagggatgt
     4021 tctcacctga gcttagaacc tttaccaaag gtgatgcgga gagatgggta agcacaacca
     4081 aaaaagccag tgattctgca ttctggcttg aggttgaagg taattccatg accgcaccaa
     4141 caggctccaa gccgagcttt cctgacggaa tgttaattct cgttgaccct gagcaggctg
     4201 ttgagccagg tgatttctgc atagccagac ttgggggtga tgagtttacc ttcaagaaac
     4261 tgatcaggga tagcggtcag gtgtttttac aaccactaaa cccacagtac ccaatgatcc
     4321 catgcaatga gagttgttcc gttgtgggga aagttatcgc tagtcagtgg cctgaagaga
     4381 cgtttggcgc tgcaaacgac gaaaactacg ctttagtagc ttaataacgc tgatagtgct
     4441 agtgtagatc gctactagag ccaggcatca aataaaacga aaggctcagt cgaaagactg
     4501 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     4561 accttcgggt gggcctttct gcgtttatat actagagtaa caccgtgcgt gttgactatt
     4621 ttacctctgg cggtgataat ggttgcttaa ttaaatgcgc acccttagcg agaggtttat
     4681 cattaaggtc aacctctgga tgttgtttcg gcatcctgca ttgaatctga gttactgtct
     4741 gttttcctga attccgatag attgtcgcac ctgattgccc gacattatcg cgggcccatt
     4801 tatacccata taaatcagca tccatgttgg aattcaggaa acccgttttt tctgacgtaa
     4861 gggtgcgcaa ctttcatgaa atccgctgaa tatttgaaca cttttagatt gagaaatctc
     4921 ggcctacctg tcatgaacaa tttgcatgac atgtctaagg cgactcgcat atctgttgaa
     4981 acacttcggt tgttaatcta tacagctgat tttcgctata ggatctacac tgtagaaaag
     5041 aaaggcccag agaagagaat gagaaccatt taccaacctt ctcgagaact taaagcctta
     5101 caaggatggg ttctacgtaa cattttagat aaactgtcgt catctccttt ttctattgga
     5161 tttgaaaagc accaatctat tttgaataat gctaccccgc atattggggc aaactttata
     5221 ctgaatattg atttggagga ttttttccca agtttaactg ctaacaaagt ttttggagtg
     5281 ttccattctc ttggttataa tcgactaata tcttcagttt tgacaaaaat atgttgttat
     5341 aaaaatctgc taccacaagg tgctccatca tcacctaaat tagctaatct aatatgttct
     5401 aaacttgatt atcgtattca gggttatgca ggtagtcggg gcttgatata tacgagatat
     5461 gccgatgacc tcaccttatc tgcacagtct atgaaaaagg ttgttaaagc acgtgatttt
     5521 ttattttcta taatcccaag tgaaggattg gttattaact caaaaaaaac ttgtattagt
     5581 gggcctcgta gtcagaggaa agttacaggt ttagttattt cacaagagaa agttgggata
     5641 ggtagagaaa aatataaaga aattagagca aagatacatc atatattttg cggtaagtct
     5701 tctgagatag aacacgttag gggatggttg tcatttattt taagtgtgga ttcaaaaagc
     5761 cataggagat taataactta tattagcaaa ttagaaaaaa aatatggaaa gaacccttta
     5821 aataaagcga agacctaagg atccggttga tattgattca gaggtataaa acgaatgagt
     5881 actgcactcg caacgctggc tgggaagctg gctgaacgtg tcggcatgga ttctgtcgac
     5941 ccacaggaac tgatcaccac tcttcgccag acggcattta aaggtgatgc cagcgatgcg
     6001 cagttcatcg cattactgat cgttgccaac cagtacggcc ttaatccgtg gacgaaagaa
     6061 atttacgcct ttcctgataa gcagaatggc atcgttccgg tggtgggcgt tgatggctgg
     6121 tcccgcatca tcaatgaaaa ccagcagttt gatggcatgg actttgagca ggacaatgaa
     6181 tcctgtacat gccggattta ccgcaaggac cgtaatcatc cgatctgcgt taccgaatgg
     6241 atggatgaat gccgccgcga accattcaaa actcgcgaag gcagagaaat cacggggccg
     6301 tggcagtcgc atcccaaacg gatgttacgt cataaagcca tgattcagtg tgcccgtctg
     6361 gccttcggat ttgctggtat ctatgacaag gatgaagccg agcgcattgt cgaaaatact
     6421 gcatacactg cagaacgtca gccggaacgc gacatcactc cggttaacga tgaaaccatg
     6481 caggagatta acactctgct gatcgccctg gataaaacat gggatgacga cttattgccg
     6541 ctctgttccc agatatttcg ccgcgacatt cgtgcatcgt cagaactgac acaggccgaa
     6601 gcagtaaaag ctcttggatt cctgaaacag aaagccgcag agcagaaggt ggcagcatga
     6661 acgcgtgcta gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     6721 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct
     6781 aggcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     7141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7561 cgaaaactca cgttaaggga ttttggtcat gactagtgct tggattctca ccaataaaaa
     7621 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     7681 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
     7741 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acagacggca
     7801 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     7861 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     7921 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     7981 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     8041 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     8101 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     8161 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     8221 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     8281 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     8341 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     8401 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     8461 gaacctctta cgtgccgatc aacgtctcat tttcgccaga tatcgacgt
//
