LOCUS       40924_40092        7109 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Recombinase expression vector pSH66, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7109)
  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 7109)
  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 7109)
  AUTHORS   Hegemann JH, Heick SB.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2010) Lehrstuhl fur Funktionelle Genomforschung 
            der Mikroorganismen, Heinrich-Heine-Universitaet, 
            Universitatsstrasse 1, Duesseldorf, NRW 40225, Germany
REFERENCE   4  (bases 1 to 7109)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7109)
  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: "Methods 
            Mol. Biol. 765, 189-206 (2011)"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (15-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..7109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            241..1360
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     rep_origin      complement(1481..1936)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2081..2097
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2107..2125
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2144..2391)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(2713..3741)
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(3839..3855)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3870..4311)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        complement(4335..4353)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4374..4390)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4398..4414)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4422..4452)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4467..4488)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4776..5364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5538..6395)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6396..6500)
                     /label=AmpR promoter
     misc_feature    6537..7040
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgcgtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatagg tctagagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc
      241 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
      301 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      361 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
      421 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      481 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      541 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt accactcttg
      601 acgacacggc ttaccggtac cgcaccagtg tcccggggga cgccgaggcc atcgaggcac
      661 tggatgggtc cttcaccacc gacaccgtct tccgcgtcac cgccaccggg gacggcttca
      721 ccctgcggga ggtgccggtg gacccgcccc tgaccaaggt gttccccgac gacgaatcgg
      781 acgacgaatc ggacgacggg gaggacggcg acccggactc ccggacgttc gtcgcgtacg
      841 gggacgacgg cgacctggcg ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc
      901 tgaccgtcga ggacatcgag gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt
      961 tgatggggct cgcgacggag ttcgcccgcg agcggggcgc cgggcacctc tggctggagg
     1021 tcaccaacgt caacgcaccg gcgatccacg cgtaccggcg gatggggttc accctctgcg
     1081 gcctggacac cgccctgtac gacggcaccg cctcggacgg cgagcaggcg ctctacatga
     1141 gcatgccctg cccctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc
     1201 atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta
     1261 tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat
     1321 atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg
     1381 ccgccatcca gtgtcgaaaa cgagctctcg agaaccctta atcggatcta tgcggtgtga
     1441 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1501 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1561 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1621 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1681 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1741 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1801 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1861 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1921 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1981 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2041 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     2101 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg gccgcaaatt aaagccttcg
     2161 agcgtcccaa aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc
     2221 tgtacagaaa aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta
     2281 taaaaaaata aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag
     2341 cggatgtggg gggagggcgt gaatgtaagc gtgacataac taattacatg actcgaggtc
     2401 gacaattcca accttaccca agagttcgcc aaactcagac atcactttan gcaaaaccgc
     2461 gccgtgcttc ttcctcggtg gcattcatca cgaaatgttc agcactacgc atacttttga
     2521 caggaaacgc aacggatatt gagtcaatat caggcattct atcgctcagc tttacagtga
     2581 caatgacggc tggcgactga atattagtgc ttacagacag cactacatat tttccgtcga
     2641 tgttgaaatc ctttctcata tgtcaccata aatatcaaat aattatagca atcatttacg
     2701 cgttaatggc taatcgccat cttccagcag gcgcaccatt gcccctgttt cactatccag
     2761 ggtacggata tagttcatga caatatttac attggtccag ccaccagctt gcatgatctc
     2821 cggtattgaa actccagcgc gggccatatc tcgcgcggct ccgacacggg cactgtgtcc
     2881 agaccaggcc aggtatctct gaccagagtc atccttagcg ccgtaaatca atcgatgagt
     2941 tgcttcaaaa atcccttcca gggcgcgagt tgatagctgg ctggtggcag atggcgcggc
     3001 aacaccattt tttctgaccc ggcaaaacag gtagttattc ggatcatcag ctacaccaga
     3061 gacggaaatc catcgctcga ccagtttagt tacccccagg ctaagtgcct tctctacacc
     3121 tgcggtgcta accagcgttt tcgttctgcc aatatggatt aacattctcc caccgtcagt
     3181 acgtgagata tctttaaccc tgatcctggc aatttcggct atacgtaaca gggtgttata
     3241 agcaatcccc agaaatgcca gattacgtat atcctggcag cgatcgctat tttccatgag
     3301 tgaacgaacc tggtcgaaat cagtgcgttc gaacgctaga gcctgttttg cacgttcacc
     3361 ggcatcaacg ttttcttttc ggatccgccg cataaccagt gaaacagcat tgctgtcact
     3421 tggtcgtggc agcccggacc gacgatgaag catgtttagc tggcccaaat gttgctggat
     3481 agtttttact gccagaccgc gcgcctgaag atatagaaga taatcgcgaa catcttcagg
     3541 ttctgcggga aaccatttcc ggttattcaa cttgcaccat gccgcccacg accggcaaac
     3601 ggacagaagc attttccagg tatgctcaga aaacgcctgg cgatccctga acatgtccat
     3661 caggttcttg cgaacctcat cactcgttgc atcgaccggt aatgcaggca aattttggtg
     3721 tacggtcagt aaattggaca tttaacactc agataatggt tttaagtaaa gtgtacagga
     3781 tcggctctgc ccctcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga
     3841 tccactagtt ctagaatccg gggttttttc tccttgacgt taaagtatag aggtatatta
     3901 acaatttttt gttgatactt ttattacatt tgaataagaa gtaatacaaa ccgaaaatgt
     3961 tgaaagtatt agttaaagtg gttatgcagt ttttgcattt atatatctgt taatagatca
     4021 aaaatcatcg cttcgctgat taattacccc agaaataagg ctaaaaaact aatcgcatta
     4081 tcatcctatg gttgttaatt tgattcgttc atttgaaggt ttgtggggcc aggttactgc
     4141 caatttttcc tcttcataac cataaaagct agtattgtag aatctttatt gttcggagca
     4201 gtgcggcgcg aggcacatct gcgtttcagg aacgcgaccg gtgaagacga ggacgcacgg
     4261 aggagagtct tccttcggag ggctgtcacc cgctcggcgg cttctaatcc gtactagagc
     4321 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4381 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc
     4441 ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc
     4501 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4561 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4621 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4681 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4741 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4801 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4861 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4921 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4981 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5041 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5101 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5161 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5221 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5281 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5341 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5401 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5461 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5521 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5581 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5641 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5701 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5761 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5821 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5881 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5941 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6001 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6061 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6121 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6181 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6241 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6301 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6361 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6421 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6481 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc cttttcatca
     6541 cgtgctataa aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag
     6601 aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaagactc
     6661 tagggggatc gccaacaaat actacctttt atcttgctct tcctgctctc aggtattaat
     6721 gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg tgattttaca
     6781 ttttacttat cgttaatcga atgtatatct atttaatctg cttttcttgt ctaataaata
     6841 tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct ttgtttattt ttttttcttc
     6901 attccgtaac tcttctacct tctttattta ctttctaaaa tccaaataca aaacataaaa
     6961 ataaataaac acagagtaaa ttcccaaatt attccatcat taaaagatac gaggcgcgtg
     7021 taagttacag gcaagcgatc cgtcctaaga aaccattatt atcatgacat taacctataa
     7081 aaataggcgt atcacgaggc cctttcgtc
//
