LOCUS       40924_45513        3772 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PCR template vector pUG74, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3772)
  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 3772)
  AUTHORS   Hegemann JH, Heick SB.
  TITLE     Delete and Repeat: A Comprehensive Toolkit for Sequential Gene 
            Knockout in the Budding Yeast Saccharomyces cerevisiae
  JOURNAL   Methods Mol. Biol. 765, 189-206 (2011)
  PUBMED    21815094
REFERENCE   3  (bases 1 to 3772)
  AUTHORS   Hegemann JH, Heick SB.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-OCT-2010) Lehrstuhl fur Funktionelle Genomforschung 
            der Mikroorganismen, Heinrich-Heine-Universitaet, 
            Universitatsstrasse 1, Duesseldorf, NRW 40225, Germany
REFERENCE   4  (bases 1 to 3772)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3772)
  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: "Methods 
            Mol. Biol. 765, 189-206 (2011)"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (13-OCT-2010) Lehrstuhl fur Funktionelle Genomforschung der 
            Mikroorganismen, Heinrich-Heine-Universitaet, Universitatsstrasse 1,
            Duesseldorf, NRW 40225, Germany"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3772
                     /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..1260
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     misc_feature    1323..1356
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1323..1356)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(1410..1428)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1686..2274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2448..3305)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3306..3410)
                     /label=AmpR promoter
     promoter        3756..3772
                     /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 accactcttg acgacacggc ttaccggtac cgcaccagtg tcccggggga
      541 cgccgaggcc atcgaggcac tggatgggtc cttcaccacc gacaccgtct tccgcgtcac
      601 cgccaccggg gacggcttca ccctgcggga ggtgccggtg gacccgcccc tgaccaaggt
      661 gttccccgac gacgaatcgg acgacgaatc ggacgacggg gaggacggcg acccggactc
      721 ccggacgttc gtcgcgtacg gggacgacgg cgacctggcg ggcttcgtgg tcgtctcgta
      781 ctccggctgg aaccgccggc tgaccgtcga ggacatcgag gtcgccccgg agcaccgggg
      841 gcacggggtc gggcgcgcgt tgatggggct cgcgacggag ttcgcccgcg agcggggcgc
      901 cgggcacctc tggctggagg tcaccaacgt caacgcaccg gcgatccacg cgtaccggcg
      961 gatggggttc accctctgcg gcctggacac cgccctgtac gacggcaccg cctcggacgg
     1021 cgagcaggcg ctctacatga gcatgccctg cccctaatca gtactgacaa taaaaagatt
     1081 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     1141 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     1201 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     1261 ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctctcg agaaccctta
     1321 atataacttc gtataatgta tgctatacga agttattagg tgatatcaga tccactagtg
     1381 gcctatgcgg ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc
     1441 caggttaacc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1501 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1561 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1621 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1681 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1741 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1801 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1861 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1921 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1981 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2041 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2101 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2161 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2221 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2281 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2341 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2461 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2521 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2581 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2641 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2701 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2761 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2821 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2881 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2941 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3001 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3061 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3421 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     3481 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     3541 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     3601 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     3661 tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac
     3721 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta ta
//
