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"
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
      241 gcgtatcacg aggccctttc gtcttcgaat aaatacctgt gacggaagat cacttcgcag
      301 aataaataaa tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa
      361 aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
      421 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
      481 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
      541 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
      601 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
      661 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
      721 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
      781 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
      841 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
      901 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
      961 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     1021 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     1081 gaatgcttaa tgaattacaa cagtttttat gcatgcattt acgttgacac catcgaatgg
     1141 tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     1201 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     1261 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     1321 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     1381 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     1441 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     1501 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     1561 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     1621 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     1681 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     1741 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     1801 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     1861 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     1921 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     1981 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     2041 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     2101 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     2161 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     2221 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     2281 cagtgagcgc aacgcaatta atgtgagtta gcgcgaattg atctggtttg acagcttatc
     2341 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     2401 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     2461 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     2521 acaattaatc atccggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
     2581 ggaaacagac catatgctcg aggactacaa agaccatgac ggtgattata aagatcatga
     2641 tatcgactac aaagatgacg acgataaacc cgggcaccca gaaacgctgg tgaaagtaaa
     2701 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2761 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2821 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2881 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2941 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3001 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3061 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3121 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3181 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3241 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3301 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3361 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3421 tagacagatc gctgagatag gtgcctcact gattaagcat tggtctagat aactgcagta
     3481 agggcgaatt ccagcacact ggcggccgtt actagtggat ccgagagctc caattcgccc
     3541 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     3601 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3661 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3721 tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3781 cattttttaa ccaataggcc gactgcgatg agtggcaggg cggggcgtaa tttttttaag
     3841 gcagttattg gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat
     3901 gaatggcaga aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa
     3961 atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc
     4021 gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt
     4081 gacgatatga tcatttattc tgcctcccag agcctgataa aaacggtgaa tccgttagcg
     4141 aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg
     4201 cggggaggca gacaaggtat agggcggcga ggcggctaca gccgatagtc tggaacagcg
     4261 cacttacggg ttgctgcgca acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc
     4321 ggctccaacg gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc
     4381 tgcccgcgcc gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc
     4441 ggaatgccgg gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc
     4501 ggatacaggg tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg
     4561 tcgtgatcaa ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg
     4621 ccccacgcca cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc
     4681 acgacggaga tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa
     4741 gaacgtccga tgagcttgga aagtgtcttc tggctgacca ccacggcgtt ctggtggccc
     4801 atctgcgcca cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg
     4861 gcccacgcct catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct
     4921 tgaatgccga tttctctgga ctgcgtggcc atgcttatct ccatgcggta gggtgccgca
     4981 cggttgcggc accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg
     5041 ttgcgtaaaa gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg
     5101 ggggtgccgc aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt
     5161 tcgcatttcg ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc
     5221 ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga
     5281 tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact
     5341 cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc cttcgactgc ccccactcgc
     5401 ataggcttgg gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc
     5461 cttgacccgc cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg
     5521 aggcttttcc ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt
     5581 ggagccggtg ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc
     5641 tcgtgggcag gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg
     5701 aagcgagcca gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg
     5761 agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc
     5821 gtaggcggtc acgactttgc gaagcaaagt ctagtgagta tactcaagca ttgagtggcc
     5881 cgccggaggc accgccttgc gctgcccccg tcgagccggt tggacaccaa aagggagggg
     5941 caggcatggc ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg
     6001 cggccagtct caagcacgcc taccgcgagc gcgagacgcc caacgctgac gccagcagga
     6061 cgccagagaa cgagcactgg gcggccagca gcaccgatga agcgatgggc cgactgcgcg
     6121 agttgctgcc agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg
     6181 ccagcccgga atggtggaag tcggccagcc aagaacagca ggcggcgttc ttcgagaagg
     6241 cgcacaagtg gctggcggac aagtacgggg cggatcgcat cgtgacggcc agcatccacc
     6301 gtgacgaaac cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc
     6361 tgtcggccaa ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag accacgtttg
     6421 cggccgctgt ggccgatcta gggctgcaac ggggcatcga gggcagcaag gcacgtcaca
     6481 cgcgcattca ggcgttctac gaggccctgg agcggccacc agtgggccac gtcaccatca
     6541 gcccgcaagc ggtcgagcca cgcgcctatg caccgcaggg attggccgaa aagctgggaa
     6601 tctcaaagcg cgttgagacg ccggaagccg tggccgaccg gctgacaaaa gcggttcggc
     6661 aggggtatga gcctgcccta caggccgccg caggagcgcg tgagatgcgc aagaaggccg
     6721 atcaagccca agagacggcc cgag
//
