pFlpAB-4 vector (V006222)

Basic Vector Information

Vector Name:
pFlpAB-4
Length:
10142 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Barrett AR, Kang Y, Inamasu KS, Son MS, Vukovich JM, Hoang TT.

pFlpAB-4 vector Map

pFlpAB-410142 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000pRO1600 oriVpRO1600 RepM13 fwdregulatoryPhenylalanine--tRNA ligase alpha subunitFLPRBSlambda repressor (ts)M13 revlac operatorlac promoterCAP binding siteoriToritonB terminatorTelAKilARBSregulatory

pFlpAB-4 vector Sequence

LOCUS       V006222                10142 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006222
VERSION     V006222
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10142)
  AUTHORS   Barrett AR, Kang Y, Inamasu KS, Son MS, Vukovich JM, Hoang TT.
  TITLE     Genetic tools for allelic replacement in Burkholderia species
  JOURNAL   Appl. Environ. Microbiol. 74 (14), 4498-4508 (2008)
   PUBMED   18502918
REFERENCE   2  (bases 1 to 10142)
  AUTHORS   Barrett AR, Kang Y, Vukovich JM, Son MS, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-DEC-2007) Microbiology, University of Hawaii at Manoa,
            2538 McCarthy Mall Snyder Hall Rm. 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 10142)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10142)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "14";
            pages: "4498-4508"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (03-DEC-2007) Microbiology, University of Hawaii at Manoa, 2538
            McCarthy Mall Snyder Hall Rm. 310, Honolulu, HI 96822, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(232..583)
                     /direction=LEFT
                     /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..1427
                     /label="pRO1600 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     primer_bind     1591..1607
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      1662..1703
                     /regulatory_class="promoter"
     CDS             1739..2749
                     /gene="pheS"
                     /label="Phenylalanine--tRNA ligase alpha subunit"
                     /note="Phenylalanine--tRNA ligase alpha subunit from
                     Burkholderia pseudomallei (strain 1710b). Accession#:
                     Q3JT08"
     CDS             complement(2964..4232)
                     /label="FLP"
                     /note="site-specific recombinase"
     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..5043
                     /label="lambda repressor (ts)"
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     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    complement(5428..5449)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            5833..5941
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     terminator      6926..6957
                     /label="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     CDS             complement(6975..7928)
                     /codon_start=1
                     /gene="telB"
                     /product="TelB"
                     /label="telB"
                     /note="confers tellurite resistance"
                     /protein_id="ABY57988.1"
                     /translation="MNATNTDVFAQVGGLEARGAKMKKRGTRFLIAALAVLAIAGIGAV
                     TGWAISPSATPGSIDVPQVLASTFSDQVPGSEGGGLGGGLPFTSAVGAFTDFMAGPAIF
                     TLGILGIVVAGAVLVFGGEFCGFVRSVCMMVIAVSMIFVSSNLVKGILGGDHDAGPAEP
                     SPRARFMAAVEAKDFARVQELIEARGAKSAADYVLAQLAVAEGLDRKPGARVVVGKAAG
                     SMAMPPAALGFTPRGEAAYAIERSAYGEPRSSIAKQYQQEWNRKAATWWAMAGVAGIIG
                     AILAAAATGFVGLAVSIRNRVKRVRDLLVMEPGAEP"
     gene            complement(6975..7928)
                     /gene="telB"
                     /label="telB"
     CDS             complement(7925..9061)
                     /codon_start=1
                     /gene="telA"
                     /product="TelA"
                     /label="telA"
                     /note="confers tellurite resistance"
                     /protein_id="ABY57989.1"
                     /translation="MNALKTTHDAKAPIVAFDMTPATLRELGLQESDVPEVHAVAQRIE
                     VGSPQTVAEFGRDVAEHTSRYADSLLDQVRNSDLDEAGEKLTQVVAKARSLNVGPLSDN
                     RSRLPLIGPLIDRFRVRSTGFMARFDTTREQIEHLVSEVQTTQQGIAQRNASLDEMFAA
                     VREEHRLLGVHIAAGKVRLAELREQAEGLRGNVGNDPGRVQELADLDAMVANLDKRIGD
                     LIALQHSAMQSLPTIRMIQANNQMLVDKFHTIREITVPAWKRQFMLALSLNEQKNAVEL
                     ATAIDDTTNDLMKRNAALLHRTSVETAKENQRLVIDVDTLKQVQTTLIKTVEDVIRIQQ
                     EGVQKRKDAEKQIAAMRGDLQAKLTRQPVRELAQQESV"
     gene            complement(7925..9061)
                     /gene="telA"
                     /label="telA"
     CDS             complement(9079..9852)
                     /codon_start=1
                     /gene="kilA"
                     /product="KilA"
                     /label="kilA"
                     /note="confers tellurite resistance"
                     /protein_id="ABY57990.1"
                     /translation="MEEQSVNMARLKGEVLPALFASPATIGEYGAGIDGADSLNELSNL
                     MEHGAVAALADKISQIVAKLADADPRKIAEKPTWFEKMLGREVERQVRYQVARKTLDQL
                     LDEAEGVAQRVRDTLRALDDMLNTHEAEVDRLRAYIQAGREFLDENPEAGAAKAGVIEF
                     DKPRERFARKLANLATLMASHEMSVTQMKLTRAQAVDMLDRFSETASVLVPVWRQHTLA
                     LITTKNMNPAMVAEAAKAHQALMRSLSQSLEGINQ"
     gene            complement(9079..9852)
                     /gene="kilA"
                     /label="kilA"
     RBS             complement(9859..9881)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(9892..9933)
                     /regulatory_class="promoter"

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