LOCUS       Exported                7648 bp ds-DNA    circular SYN 09-1-2013
DEFINITION  BAC vector with inducible copy number, suitable for cloning toxic 
            inserts.
ACCESSION   EU101022
VERSION     .
KEYWORDS    pSMART BAC v2.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7648)
  AUTHORS   Lucigen
  TITLE     Direct Submission
COMMENT     This sequence does not include a 3.3-kb lacZ-sacB stuffer fragment 
            inserted at the origin. 
            
            Linearization with an enzyme such as BamHI, HindIII, or EcoRI 
            removes the stuffer.
FEATURES             Location/Qualifiers
     source          1..7648
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        complement(61..79)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      102..147
                     /note="rrnBT1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        151..253
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene"
     CDS             254..913
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      927..958
                     /note="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    complement(984..1017)
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1035..1433
                     /note="cos"
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     misc_feature    1694..2167
                     /gene="sopC"
                     /note="sopC"
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(2240..3211)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /note="sopB"
                     /note="
                     "
                     /protein_id="
                     "
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(3211..4386)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /note="sopA"
                     /note="
                     "
                     /protein_id="
                     "
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    4712..4962
                     /gene="incC"
                     /note="incC"
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(4965..5720)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /note="repE"
                     /note="
                     "
                     /protein_id="
                     "
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      5811..6030
                     /note="ori2"
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      6106..6720
                     /note="oriV"
                     /note="origin of replication for the bacterial F plasmid"
     terminator      7487..7514
                     /note="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     promoter        7573..7591
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"