LOCUS       40924_26151        3988 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PCR template vector pJJH726, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3988)
  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   2  (bases 1 to 3988)
  AUTHORS   Gueldener U, Heinisch JH, Voss D, Koehler G, Hegemann JH.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany
REFERENCE   3  (bases 1 to 3988)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3988)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2002"; volume: "30"; issue: "6"; pages: "E23"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3988
                     /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)."
     CDS             complement(220..1023)
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine-5'-phosphate decarboxylase"
                     /label=URA3
                     /note="from Kluyveromyces lactis"
                     /protein_id="AAG34531.1"
                     /translation="MSTKSYTSRAETHASPVASKLLRLMDEKKTNLCASLDVRSTDELL
                     KLVETLGPYICLLKTHVDILDDFSYEGTVVPLKALAEKYKFLIFEDRKFADIGNTVKLQ
                     YTSGVYRIAEWSDITNAHGVTGAGIVAGLKQGAQEVTKEPRGLLMLAELSSKGSLAHGE
                     YTKGTVDIAKSDKDFVIGFIAQNDMGGREEGFDWLIMTPGVGLDDKGDALGQQYRTVDE
                     VVSGGSDIIIVGRGLFAKGRDPKVEGERYRNAGWEAYQKRISAPH"
     gene            complement(220..1023)
                     /gene="URA3"
                     /label=URA3
     misc_feature    1523..1576
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1539..1572)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(1626..1644)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1902..2490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2664..3521)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3522..3626)
                     /label=AmpR promoter
     promoter        3972..3988
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc ccaatacaac agatcacgtg
      121 atcttttgta agatgaagtt gaagtgagtg ttgcaccgtg ccaatgcagg tggctattag
      181 attaaatatg tgatttgttc tattaagttt cctgtataat taatggggag cgctgattct
      241 cttttggtac gcttcccatc cagcatttct gtatctttca ccttcaacct taggatctct
      301 acccttggcg aaaagtcctc tgccaacaat gatgatatct gatccaccac ttacaacttc
      361 gtcgacggtt ctgtactgct gacccaatgc atcgcctttg tcgtctaaac ctacacctgg
      421 ggtcatgatt agccaatcaa acccttcttc tcttcctccc atatcgttct gagcaatgaa
      481 cccaataacg aaatctttat cactctttgc aatatcaacg gtacccttag tatattcacc
      541 gtgtgctaga gaacccttgg aagacaattc agcaagcatc aataatcccc ttggttcttt
      601 ggtgacctct tgcgcacctt gtttcaagcc agcaacaata ccagcaccag taaccccgtg
      661 ggcgttggtg atatcagacc attctgcgat acggtaaacg cccgatgtat attgtaattt
      721 gactgtgtta ccgatatcgg cgaattttct gtcctcaaat atcaagaact tgtatttctc
      781 tgccaatgct ttcaatggaa cgacagtacc ctcataactg aaatcatcca agatatcaac
      841 gtgtgttttc aaaaggcaaa tgtatggacc caacgtttca acaagtttca atagctcatc
      901 agtcgaacga acgtcaagag aagcacacaa attggtcttc ttttcatcca ttaaacgtaa
      961 aagtttcgat gcaaccggac ttgcatgagt ctcagctcta ctggtatatg attttgtgga
     1021 catggtgcaa ctaattgacg ggagtgtatt gacgctggcg tactggcttt cacaaaatgg
     1081 cccaatcaca accacatctt agatagttga aatgacttta gataacatca attgagatga
     1141 gcttaatcat gtcaaagcta aaagtgtcac catgaacgac aattcttaag caaatcacgt
     1201 gatatagatc cacgaataac caccatttga tgctcgaggc aagtaatgtg tgtaaaaaaa
     1261 tgcgttacca ccatccaatg cagaccgatc ttctacccag aatcacatat atttatgtac
     1321 cgagtacctt ttttctatct tccaattgct tctcccatat gattgtctcc gtaagctcga
     1381 aatttctaag ttggatttta atcttcacgc aggatgacag ttcgatgagc ttctgaggag
     1441 tgtttagaac ataatcagtt tatccatggt ctatctcttc ttgtcgcttt ttctcctcga
     1501 tagaacctaa ataaaacgag ctctcgagaa cccttaatat aacttcgtat aatgtatgct
     1561 atacgaagtt attaggtgat atcagatcca ctagtggcct atgcggccgc ggatctgccg
     1621 gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc
     1681 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     2101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2341 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2581 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2641 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2701 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2761 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2821 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2881 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2941 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3001 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3061 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3121 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3181 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3241 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3301 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3361 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3421 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3481 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3541 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3601 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3661 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3721 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3781 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3841 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     3901 caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc
     3961 atacacatac gatttaggtg acactata
//
