LOCUS       Exported                3446 bp ds-DNA     circular SYN 25-MAR-2021
DEFINITION  Host strain: CC118 Lambda pir.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3446)
  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;98:15264-9
  PUBMED    11742086
REFERENCE   2  (bases 1 to 3446)
  AUTHORS   Lionello Bossi
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 29, 2021 from SnapGene 5.2.4
            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"; volume: "98"; pages: "15264-9"
FEATURES             Location/Qualifiers
     source          1..3446
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     CDS             133..993
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=ßla (AmpR)
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1492..1848
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             1913..1936
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     1913..1933
                     /label=Fw primer annealing seq
     protein_bind    complement(1962..1995)
                     /label=FRT-1
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated recombination"
     CDS             2370..3164
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=aph (KanR)
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(3355..3388)
                     /label=FRT-2
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated recombination"
     primer_bind     complement(3399..3418)
                     /label=Rv primer annealing seq
ORIGIN
        1 gcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
       61 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      121 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      181 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      241 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      301 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      361 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      421 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      481 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
      541 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
      601 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
      661 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
      721 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
      781 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
      841 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
      901 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
      961 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1021 ctttagattg atttaaaact tcatttttaa ttttggtgca ctctcagtac aatctgctct
     1081 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     1141 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     1201 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     1261 catcaccgaa acgcgcgagg cagcaaggag atggcgccca acagtccccc ggccacgggg
     1321 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     1381 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     1441 ccggccacga tgcgtccggc gtagaggatc cagcagttca acctgttgat agtacgtact
     1501 aagctctcat gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa
     1561 cgaactaaac cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat
     1621 gtttcacgta ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa
     1681 tagcctctaa ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagctt
     1741 ttaaggttta acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga
     1801 gcctctcaaa gcaattttca gtgacacagg aacacttaac ggctgacatg gccttagaag
     1861 gcgttttaga agccctaatg gatggagtca aaacaaacta gcgatcgaat tcgactacaa
     1921 agatgacgac gataaatagt aatgtaggct ggagctgctt cgaagttcct atactttcta
     1981 gagaatagga acttcggaat aggaacttca agatcccctc acgctgccgc aagcactcag
     2041 ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct
     2101 gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga
     2161 gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga
     2221 cagcaagcga accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca
     2281 aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg
     2341 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     2401 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     2461 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     2521 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     2581 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     2641 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     2701 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     2761 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     2821 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2881 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     2941 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3001 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3061 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3121 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3181 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3241 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3301 gcgcggggat ctcatgctgg agttcttcgc ccaccccagc ttcaaaagcg ctctgaagtt
     3361 cctatacttt ctagagaata ggaacttcgg aataggaact aaggaggata ttcatatgcg
     3421 gaattcccgg gagagctcga tgatct
//
