LOCUS       Exported               10508 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBACr-AraT7940_rbs9, complete sequence.
ACCESSION   JQ394800
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10508)
  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 10508)
  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 10508)
  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 10508)
  AUTHORS   Moser F, Voigt CA.
  TITLE     Genetic Circuit Performance under Conditions Relevant for Industrial
            Bioreactors
  JOURNAL   Unpublished
REFERENCE   5  (bases 1 to 10508)
  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 10508)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10508
                     /organism="Cloning vector pBACr-AraT7940_rbs9"
                     /mol_type="other DNA"
                     /label=derived from Escherichia coli strain DH10b
                     /note="derived from Escherichia coli strain DH10b"
                     /db_xref="taxon:1175532"
     CDS             complement(102..980)
                     /codon_start=1
                     /gene="araC"
                     /product="araC"
                     /label=araC
                     /protein_id="AFI26418.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(102..980)
                     /gene="araC"
                     /label=araC
     promoter        1007..1291
                     /gene="araBAD"
                     /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)"
     regulatory      1008..1296
                     /regulatory_class="promoter"
                     /note="PBad"
     regulatory      1302..1334
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS9"
     misc_feature    1335..4001
                     /gene="T7ptag"
                     /label=similar to T7ptag amberstop
                     /note="similar to T7ptag amberstop"
     gene            1335..4001
                     /gene="T7ptag"
                     /label=T7ptag
     misc_feature    1350..1352
                     /gene="T7ptag"
                     /label=amberstop
                     /note="amberstop"
     misc_feature    1668..1670
                     /gene="T7ptag"
                     /label=amberstop
                     /note="amberstop"
     CDS             4021..4050
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4066..4083
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      4189..4346
                     /regulatory_class="terminator"
                     /note="Trrn"
     terminator      4309..4355
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      4411..4799
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    4483..4830
                     /label=R6K
                     /note="R6K"
     CDS             complement(4849..5643)
                     /codon_start=1
                     /gene="KanR"
                     /product="KanR"
                     /label=KanR
                     /protein_id="AFI26420.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(4849..5643)
                     /gene="KanR"
                     /label=KanR
     rep_origin      6049..6268
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             6359..7114
                     /codon_start=1
                     /gene="repE"
                     /product="repE"
                     /label=repE
                     /protein_id="AFI26416.1"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     gene            6359..7114
                     /gene="repE"
                     /label=repE
     misc_feature    7117..7367
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             7693..8868
                     /codon_start=1
                     /gene="parA"
                     /product="parA"
                     /label=parA
                     /protein_id="AFI26419.1"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     gene            7693..8868
                     /gene="parA"
                     /label=parA
     CDS             8868..9839
                     /codon_start=1
                     /gene="parB"
                     /product="parB"
                     /label=parB
                     /protein_id="AFI26417.1"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     gene            8868..9839
                     /gene="parB"
                     /label=parB
     gene            9912..10429
                     /gene="parC"
                     /label=parC
     misc_feature    9912..10385
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"