LOCUS       40924_45503        4009 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PCR template vector pUG6, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4009)
  AUTHORS   Guldener U, Heck S, Fielder T, Beinhauer J, Hegemann JH.
  TITLE     A new efficient gene disruption cassette for repeated use in budding
            yeast
  JOURNAL   Nucleic Acids Res. 24 (13), 2519-2524 (1996)
  PUBMED    8692690
REFERENCE   2  (bases 1 to 4009)
  AUTHORS   Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH.
  TITLE     A second set of loxP marker cassettes for Cre-mediated multiple gene
            knockouts in budding yeast
  JOURNAL   Nucleic Acids Res. 30 (6), E23 (2002)
  PUBMED    11884642
REFERENCE   3  (bases 1 to 4009)
  AUTHORS   Gueldener U, Hegemann JH, Heck S, Fiedler T, Beinhauer JD.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany
REFERENCE   4  (bases 1 to 4009)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4009)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "1996"; volume: "24"; issue: "13"; pages: 
            "2519-2524"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2002"; volume: "30"; issue: "6"; pages: "E23"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..86
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            141..1497
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     misc_feature    1560..1593
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1560..1593)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(1647..1665)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1923..2511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2685..3542)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3543..3647)
                     /label=AmpR promoter
     promoter        3993..4009
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc tgtttagctt gcctcgtccc
      121 cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg
      181 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
      241 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct
      301 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc
      361 acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc
      421 atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac
      481 aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca acatggatgc
      541 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
      601 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
      661 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
      721 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
      781 ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
      841 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
      901 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
      961 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     1021 aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     1081 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     1141 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     1201 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     1261 gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa aaagattctt
     1321 gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca
     1381 aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta
     1441 agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg
     1501 tcgattcgat actaacgccg ccatccagtg tcgaaaacga gctctcgaga acccttaata
     1561 taacttcgta taatgtatgc tatacgaagt tattaggtga tatcagatcc actagtggcc
     1621 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     1681 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1741 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1801 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1861 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1921 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1981 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2041 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2101 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2161 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2221 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2281 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2341 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     2401 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2461 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2521 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2581 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2641 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2701 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2761 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2821 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2881 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2941 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3001 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3061 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3121 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3181 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3241 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3301 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3361 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3421 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3481 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3541 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3601 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3661 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3721 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3781 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3841 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     3901 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     3961 taatacataa ccttatgtat catacacata cgatttaggt gacactata
//
