LOCUS       40924_7026        5566 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBR939b, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5566)
  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 5566)
  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 5566)
  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 5566)
  AUTHORS   Moser F, Voigt CA.
  TITLE     Genetic Circuit Performance under Conditions Relevant for Industrial
            Bioreactors
  JOURNAL   Unpublished
REFERENCE   5  (bases 1 to 5566)
  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 5566)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 5566)
  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            "Unpublished"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..5566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        211..229
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             296..1009
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1074..1168
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1225..1254
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             1270..1287
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1513..1559
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        1685..1789
                     /label=AmpR promoter
     CDS             1790..2647
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2820..3408
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3594..3734)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3839..4027)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gcggccgcat tacgctgact tgacgggacg gcggctttgt tactagtcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
      181 agccagccgg aagggccgat cccgcgaaat taatacgact cactataggg agaccacaac
      241 ggtttccctc tagacctaac taactaaaga ttaactttat aaggaggaaa aacatatgag
      301 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
      361 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
      421 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac
      481 tactttgact tatggtgttc aatgcttttc aagataccca gatcatatga aacggcatga
      541 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
      601 tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag
      661 aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga
      721 atacaactat aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa
      781 agttaacttc aaaattagac acaacattga agatggaagc gttcaactag cagaccatta
      841 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
      901 cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga
      961 gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaaa ggcctgcagc
     1021 aaacgacgaa aactacgctt tagtagctta ataagcttaa ttagctgagc ttggactcct
     1081 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     1141 ccgccgggcg ttttttattg gtgagaatcc aagctagctt ggcgagattt tcaggagcta
     1201 aggatcaatt cgaagcttgg gcccgaacaa aaactcatct cagaagagga tctgaatagc
     1261 gccgtcgacc atcatcatca tcatcattga gtttaaacgg tctccagctt ggctgttttg
     1321 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     1381 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     1441 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     1501 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1561 tgttgtttgt cggtgaacta attcttgaag acgaaagggc ctcgtgatac gcctattttt
     1621 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1681 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1741 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1801 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1861 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1921 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1981 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     2041 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2101 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2161 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2221 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2281 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     2341 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2401 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2461 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2521 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2581 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2641 gcattggtaa cccgggacca agtttactca tatatacttt agattgattt aaaacttcat
     2701 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2761 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2821 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2881 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2941 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3001 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3061 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3121 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3181 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3241 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3301 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3361 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3421 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3481 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3541 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3601 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     3661 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3721 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3781 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3841 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3901 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3961 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4021 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     4081 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     4141 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     4201 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     4261 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     4321 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     4381 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     4441 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     4501 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
     4561 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
     4621 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     4681 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     4741 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     4801 cggtccagtg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     4861 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     4921 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     4981 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     5041 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     5101 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     5161 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     5221 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     5281 catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     5341 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     5401 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     5461 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     5521 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatc
//
