LOCUS       sequence-334524-        2957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for generating C-terminal hemagglutinin (1xHA) 
            epitope tag fusions to bacterial genes by ""Lambda red"" 
            recombineering..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2957)
  AUTHORS   Uzzau S, Figueroa-Bossi N, Rubino S, Bossi L
  TITLE     Epitope tagging of chromosomal genes in Salmonella.
  JOURNAL   Proc Natl Acad Sci U S A. 2001 Dec 18;98(26):15264-9. doi: 
            10.1073/pnas.261348198. Epub 2001 Dec 11.
  PUBMED    11742086
REFERENCE   2  (bases 1 to 2957)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.261348198"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2001-12-18- 18"; volume: "98"; issue: "26"; pages: "15264-9"
COMMENT     Sequence Label: pSU314
FEATURES             Location/Qualifiers
     source          1..2957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(918..937)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1078..1100
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1239..1258
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      1346..1734
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             1744..1770
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1750..1776
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     protein_bind    complement(1802..1849)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1866..2525)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHRFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2440..2459
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2526..2627)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2734..2767)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        2853..2957
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
      961 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     1021 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     1081 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcaaggag
     1141 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     1201 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     1261 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     1321 cggccacgat gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta
     1381 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat
     1441 gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa cgtactaagc
     1501 tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat aaatcagcaa
     1561 cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat aaggctttta
     1621 aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca ctgagaagcc
     1681 cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct gacatgggaa
     1741 ttctatccgt atgatgttcc tgattatgct agcctctagt aatgtaggct ggagctgctt
     1801 cgaagttcct atactttcta gagaatagga acttcggaat aggaacttca tttaaatggc
     1861 gcgccttacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     1921 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     1981 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     2041 ttggccacgt ttaaatcgaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     2101 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     2161 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     2221 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     2281 agctcaccgt ctttcattgc catacgtaat tccggatgag cattcatcag gcgggcaaga
     2341 atgtgaataa aggccggata aaacctgtgc ttatttttct ttacggtctt taaaaaggcc
     2401 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     2461 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     2521 tccattttag cttccttagc tcctgaaaat ctcgacaact caaaaaatac gcccggtagt
     2581 gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcgac gtctcatttt
     2641 cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct
     2701 gcgaagtgat cttccgtcac aggtaggcgc gccgaagttc ctatactttc tagagaatag
     2761 gaacttcgga ataggaacta aggaggatat tcatatgcgg aattcccggg agagctcgat
     2821 gatctgcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2881 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2941 attgaaaaag gaagagt
//
