LOCUS       40924_45508        3580 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PCR template vector pUG66, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3580)
  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 3580)
  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 3580)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3580)
  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..3580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    54..87
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        142..485
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             486..866
                     /codon_start=1
                     /gene="ble"
                     /product="bleomycin resistance protein"
                     /label=ble
                     /protein_id="AAG34545.1"
                     /translation="MADQATPNLPSRDFDSTAAFYERLGFGIVFRDAGWMILQRGDLKL
                     EFFAHPGLDPLASWFSCCLRLDDLAEFYRQCKSVGIQETSSGYPRIHAPELQEWGGTMA
                     ALVDPDGTLLRLIQNELLAGIS"
     gene            486..866
                     /gene="ble"
                     /label=ble
     terminator      872..1069
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    1132..1165
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1132..1165)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(1219..1237)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1495..2083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2257..3114)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3115..3219)
                     /label=AmpR promoter
ORIGIN
        1 agaacgcggc cgccagctga agcttcgtac gctgcaggtc gacaaccctt aatataactt
       61 cgtataatgt atgctatacg aagttattag gtctagagat ctgtttagct tgcctcgtcc
      121 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
      181 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
      241 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
      301 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
      361 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
      421 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
      481 caaccatggc cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccacggccg
      541 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
      601 agcgcgggga tctcaagctg gagttcttcg cccaccccgg gctcgatccc ctcgcgagtt
      661 ggttcagctg ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg
      721 tcggcatcca ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt
      781 ggggaggcac gatggccgct ttggtcgacc cggacgggac gctcctgcgc ctgatacaga
      841 acgaattgct tgcaggcatc tcatgatcag tactgacaat aaaaagattc ttgttttcaa
      901 gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag
      961 cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca
     1021 gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg
     1081 atactaacgc cgccatccag tgtcgaaaac gagctctcga gaacccttaa tataacttcg
     1141 tataatgtat gctatacgaa gttattaggt gatatcagat ccactagtgg cctatgcggc
     1201 cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct
     1261 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1681 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1921 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3241 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3301 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3361 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3421 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     3481 tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat
     3541 aaccttatgt atcatacaca tacgatttag gtgacactat
//
