LOCUS       40924_33417        5027 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNOR10-20, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5027)
  AUTHORS   Anderson JC, Voigt CA, Arkin AP.
  TITLE     Environmental signal integration by a modular AND gate
  JOURNAL   Mol. Syst. Biol. 3, 133 (2007)
  PUBMED    17700541
REFERENCE   2  (bases 1 to 5027)
  AUTHORS   Salis HM, Mirsky EA, Voigt CA.
  TITLE     Automated design of synthetic ribosome binding sites to control 
            protein expression
  JOURNAL   Nat. Biotechnol. 27 (10), 946-950 (2009)
  PUBMED    19801975
REFERENCE   3  (bases 1 to 5027)
  AUTHORS   Tamsir A, Tabor JJ, Voigt CA.
  TITLE     Robust multicellular computing using genetically encoded NOR gates 
            and chemical 'wires'
  JOURNAL   Nature 469 (7329), 212-215 (2011)
  PUBMED    21150903
REFERENCE   4  (bases 1 to 5027)
  AUTHORS   Moser F, Voigt CA.
  TITLE     Genetic Circuit Performance under Conditions Relevant for Industrial
            Bioreactors
  JOURNAL   Unpublished
REFERENCE   5  (bases 1 to 5027)
  AUTHORS   Anderson JC, Moser F, Tamsir A, Salis H.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JAN-2012) Biological Engineering, MIT, 500 Tech 
            Square, Rm 209D, Cambridge, MA 02139, USA
REFERENCE   6  (bases 1 to 5027)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 5027)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Syst. 
            Biol. 3, 133 (2007)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Nat. 
            Biotechnol."; date: "2009"; volume: "27"; issue: "10"; pages: 
            "946-950"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Nature"; 
            date: "2011"; volume: "469"; issue: "7329"; pages: "212-215"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (06-JAN-2012) Biological Engineering, MIT, 500 Tech Square, Rm 209D,
            Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 6; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(78..121)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    124..145
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     promoter        145..429
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     protein_bind    458..476
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      506..511
                     /label=Ptet BBa_R0040
                     /note="Ptet BBa_R0040"
                     /regulatory_class="promoter"
     regulatory      520..530
                     /label=BBa_B0033
                     /note="BBa_B0033"
                     /regulatory_class="ribosome_binding_site"
     CDS             537..1247
                     /label=lambda repressor
                     /note="phage lambda repressor"
     CDS             1248..1280
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     terminator      1328..1399
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1415..1442
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      1455..1489
                     /label=BBa_J23117
                     /note="BBa_J23117"
                     /regulatory_class="promoter"
     regulatory      1498..1509
                     /label=BBa_B0034
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             1498..1509
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1516..2391
                     /label=araC
                     /note="L-arabinose regulatory protein"
     CDS             2421..3041
                     /label=TetR
                     /note="tetracycline repressor TetR"
     misc_feature    3048..3068
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3069..3128
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     rep_origin      complement(3375..3963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4051..4145)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4169..4825)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(4826..4928)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tcgagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
       61 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
      121 ctggaattcg cggccgcttc tagagagaaa ccaattgtcc atattgcatc agacattgcc
      181 gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa
      241 gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata
      301 atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag
      361 catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg
      421 tttctccata cccgtttttt tgggctagcg actagagtcc ctatcagtga tagagattga
      481 catccctatc agtgatagat ataatgagca ctactagagt cacacaggac tactagatga
      541 gcacaaaaaa gaaaccatta acacaagagc agcttgagga cgcacgtcgc cttaaagcaa
      601 tttatgaaaa aaagaaaaat gaacttggct tatcccagga atctgtcgca gacaagatgg
      661 ggatggggca gtcaggcgtt ggtgctttat ttaatggcat caatgcatta aatgcttata
      721 acgccgcatt gcttgcaaaa attctcaaag ttagcgttga agaatttagc ccttcaatcg
      781 ccagagaaat ctacgagatg tatgaagcgg ttagtatgca gccgtcactt agaagtgagt
      841 atgagtaccc tgttttttct catgttcagg cagggatgtt ctcacctgag cttagaacct
      901 ttaccaaagg tgatgcggag agatgggtaa gcacaaccaa aaaagccagt gattctgcat
      961 tctggcttga ggttgaaggt aattccatga ccgcaccaac aggctccaag ccaagctttc
     1021 ctgacggaat gttaattctc gttgaccctg agcaggctgt tgagccaggt gatttctgca
     1081 tagccagact tgggggtgat gagtttacct tcaagaaact gatcagggat agcggtcagg
     1141 tgtttttaca accactaaac ccacagtacc caatgatccc atgcaatgag agttgttccg
     1201 ttgtggggaa agttatcgct agtcagtggc ctgaagagac gtttggcgct gcaaacgacg
     1261 aaaactacgc tttagtagct taataacgct gatagtgcta gtgtagatcg ctactagagc
     1321 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     1381 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     1441 cgtttatact agagttgaca gctagctcag tcctagggat tgtgctagct actagagaaa
     1501 gaggagaaat actagatggc tgaagcgcaa aatgatcccc tgctgccggg atactcgttt
     1561 aatgcccatc tggtggcggg tttaacgccg attgaggcca acggttatct cgattttttt
     1621 atcgaccgac cgctgggaat gaaaggttat attctcaatc tcaccattcg cggtcagggg
     1681 gtggtgaaaa atcagggacg agaatttgtt tgccgaccgg gtgatatttt gctgttcccg
     1741 ccaggagaga ttcatcacta cggtcgtcat ccggaggctc gcgaatggta tcaccagtgg
     1801 gtttactttc gtccgcgcgc ctactggcat gaatggctta actggccgtc aatatttgcc
     1861 aatacggggt tctttcgccc ggatgaagcg caccagccgc atttcagcga cctgtttggg
     1921 caaatcatta acgccgggca aggggaaggg cgctattcgg agctgctggc gataaatctg
     1981 cttgagcaat tgttactgcg gcgcatggaa gcgattaacg agtcgctcca tccaccgatg
     2041 gataatcggg tacgcgaggc ttgtcagtac atcagcgatc acctggcaga cagcaatttt
     2101 gatatcgcca gcgtcgcaca gcatgtttgc ttgtcgccgt cgcgtctgtc acatcttttc
     2161 cgccagcagt tagggattag cgtcttaagc tggcgcgagg accaacgtat cagccaggcg
     2221 aagctgcttt tgagcaccac ccggatgcct atcgccaccg tcggtcgcaa tgttggtttt
     2281 gacgatcaac tctatttctc gcgggtattt aaaaaatgca ccggggccag cccgagcgag
     2341 ttccgtgccg gttgtgaaga aaaagtgaat gatgtagccg tcaagttgtc ataatactag
     2401 atcacacagg aaaggcctcg atgtccagat tagataaaag taaagtgatt aacagcgcat
     2461 tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc
     2521 taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg
     2581 ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa
     2641 gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc
     2701 gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg
     2761 aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac
     2821 tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg
     2881 ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta
     2941 tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca
     3001 tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc ctaataatac tagtagcggc
     3061 cgctgcagtc cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac
     3121 cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg
     3181 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatcctaggg cgttcggctg
     3241 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3301 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3361 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3421 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3481 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3541 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     3601 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3661 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3721 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3781 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3841 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3901 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3961 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4021 taagggattt tggtcatgac ttgtgcttgg attctcacca ataaaaaacg cccggcggca
     4081 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     4141 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt
     4201 gtaattcatt aagcattctg ccgacatgga agccatcaca gacggcatga tgaacctgaa
     4261 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     4321 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     4381 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     4441 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     4501 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     4561 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     4621 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     4681 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     4741 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     4801 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     4861 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     4921 gccgatcaac gtctcatttt cgccagatat cgacgtctaa gaaaccatta ttatcatgac
     4981 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacc
//
