pFlagTEM1 vector (V006234)

LOCUS       40924_20171        6744 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFlagTEM1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6744)
  AUTHORS   Manuela R, Sun Y, Wilson PR, Tran QT, Chessa D, Andrews-Polymenis 
            HL, Lawhon SD, Figueiredo JF, Tsolis RM, Adams GL, Baumler AJ.
  TITLE     Host restriction of Salmonella enterica serotype Typhi is not caused
            by functional alteration of SipA, SopB, or SopD
  JOURNAL   Infect. Immun. 73 (12), 7817-7826 (2005)
  PUBMED    16299271
REFERENCE   2  (bases 1 to 6744)
  AUTHORS   Sun YH, Rolan HG, Tsolis RM.
  TITLE     Injection of flagellin into the host cell cytosol by Salmonella 
            enterica serotype Typhimurium
  JOURNAL   J. Biol. Chem. 282 (47), 33897-33901 (2007)
  PUBMED    17911114
REFERENCE   3  (bases 1 to 6744)
  AUTHORS   Sun Y, Tsolis RM.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-JAN-2008) Medical Microbiology and Immunology, 
            UCDavis, School of Medicine, 451 Health Science Dr. GBSF Room 5415A,
            Davis, CA 95616, USA
REFERENCE   4  (bases 1 to 6744)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6744)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Infect. 
            Immun."; date: "2005"; volume: "73"; issue: "12"; pages: "7817-7826"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "J. Biol. 
            Chem."; date: "2007"; volume: "282"; issue: "47"; pages: 
            "33897-33901"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (31-JAN-2008) Medical Microbiology and Immunology, UCDavis, School 
            of Medicine, 451 Health Science Dr. GBSF Room 5415A, Davis, CA 
            95616, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        371..473
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             474..1106
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQF"
     promoter        1126..1203
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1204..2283
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        2518..2547
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    2555..2571
                     /label=lac operator
                     /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)."
     CDS             2603..2668
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2678..3463
                     /codon_start=1
                     /label=bla(M)
                     /note="beta-lactamase lacking the signal sequence"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     promoter        complement(3539..3557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3567..3583)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4293..4952)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(4953..5722)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"