LOCUS       Exported                7680 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFlp-AB7, complete sequence.
ACCESSION   FJ384987
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7680)
  AUTHORS   Norris MH, Kang Y, Barrett AR, Lu D, Hoang TT.
  TITLE     Glyphosate resistance as a novel non-antibiotic selectable marker in
            the select agent Burkholderia pseudomallei
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7680)
  AUTHORS   Norris MH, Kang Y, Barrett AR, Lu D, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-OCT-2008) Molecular Biology and Bio-Engineering, 
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, 
            Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 7680)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7680
                     /organism="Cloning vector pFlp-AB7"
                     /mol_type="other DNA"
                     /db_xref="taxon:578227"
     rep_origin      187..569
                     /label=ori1600 broad host range origin of replication
                     /note="ori1600 broad host range origin of replication"
     rep_origin      232..583
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             597..1430
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein; recognizes ori1600 for broad 
                     host range replication"
                     /protein_id="ACJ70057.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            597..1430
                     /gene="rep"
                     /label=rep
     primer_bind     1591..1607
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1662..1703
                     /regulatory_class="promoter"
                     /note="PCS12; rpsL promoter from Burkholderia pseudomallei"
     CDS             1739..2752
                     /codon_start=1
                     /gene="pheS"
                     /product="pheS"
                     /label=pheS
                     /note="mutant phenylalanine tRNA synthetase alpha subunit; 
                     incorporates phenylalanine and chlorinated phenylalanine on
                     tRNA"
                     /protein_id="ACJ70058.1"
                     /translation="MDLDQIVADAQQSFEGAADITTLENEKARFLGKSGALTELLKGLG
                     KLDPETRKTEGARINVAKQQVEAALNARRQALADALLNQRLAAEAIDVTLPGRGAGAGS
                     LHPVMRTWERVEQIFRSIGFDVADGPEIETDWYNFTALNSPENHPARSMQDTFYVDGKD
                     ADGRPLLLRTHTSPMQVRYARMNRPPIKVIAPGRTYRVDSDATHSPMFNQVEGLWIDEN
                     VSFADLKGAYTDFLKKFFERDDILVRFRPSYFPFTEPSAEIDMMFEHGKNAGKWLEISG
                     SGQVHPTVIRNMGLDPERYIGFGFGSGLERLTMLRYGVQDLRLFFENDLRFLRQFA"
     gene            1739..2752
                     /gene="pheS"
                     /label=pheS
     CDS             complement(2961..4232)
                     /codon_start=1
                     /gene="flp"
                     /product="Flp"
                     /label=flp
                     /note="FRT recombinase; promotes recombination between FRT 
                     sequences"
                     /protein_id="ACJ70059.1"
                     /translation="MPQFGILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     gene            complement(2961..4232)
                     /gene="flp"
                     /label=flp
     RBS             4236..4244
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             4333..5046
                     /codon_start=1
                     /gene="ci857"
                     /product="CI857"
                     /label=ci857
                     /note="cI repressor"
                     /protein_id="ACJ70060.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     gene            4333..5046
                     /gene="ci857"
                     /label=ci857
     primer_bind     complement(5335..5351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5359..5375
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5383..5413)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5428..5449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     oriT            5688..5948
     oriT            5833..5941
                     /note="incP origin of transfer"
     rep_origin      5970..6565
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6921..7361)
                     /codon_start=1
                     /gene="gat"
                     /product="Gat"
                     /label=gat
                     /note="glyphosate acetyl transferase; confers resistance to
                     glyphosate"
                     /protein_id="ACJ70061.1"
                     /translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
                     YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
                     MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
     gene            complement(6921..7361)
                     /gene="gat"
                     /label=gat
     RBS             7397..7419
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(7430..7471)
                     /regulatory_class="promoter"
                     /note="PCS12; rpsL promoter from Burkholderia cenocepacia"