LOCUS       40924_40107        7565 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Recombinase expression vector pSH69, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7565)
  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 7565)
  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 7565)
  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 7565)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7565)
  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..7565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            241..1816
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin
                     resistance"
     rep_origin      complement(1937..2392)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2537..2553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2563..2581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2600..2847)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(3169..4197)
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(4295..4311)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4326..4767)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        complement(4791..4809)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4830..4846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4854..4870)
                     /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(4878..4908)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4923..4944)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5232..5820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5994..6851)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6852..6956)
                     /label=AmpR promoter
     misc_feature    6993..7496
                     /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 aaaaagcctg
      601 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
      661 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
      721 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
      781 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
      841 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
      901 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcg atcgctgcgg
      961 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     1021 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     1081 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     1141 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     1201 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     1261 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     1321 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     1381 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     1441 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     1501 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     1561 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taatcagtac
     1621 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     1681 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     1741 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     1801 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagtgt cgaaaacgag
     1861 ctctcgagaa cccttaatcg gatctatgcg gtgtgaaata ccgcacagat gcgtaaggag
     1921 aaaataccgc atcaggaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     1981 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2041 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2101 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2161 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2221 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2281 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2341 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcgcgccat
     2401 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2461 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2521 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     2581 ggcgaattgg gtaccggccg caaattaaag ccttcgagcg tcccaaaacc ttctcaagca
     2641 aggttttcag tataatgtta catgcgtaca cgcgtctgta cagaaaaaaa agaaaaattt
     2701 gaaatataaa taacgttctt aatactaaca taactataaa aaaataaata gggacctaga
     2761 cttcaggttg tctaactcct tccttttcgg ttagagcgga tgtgggggga gggcgtgaat
     2821 gtaagcgtga cataactaat tacatgactc gaggtcgaca attccaacct tacccaagag
     2881 ttcgccaaac tcagacatca ctttangcaa aaccgcgccg tgcttcttcc tcggtggcat
     2941 tcatcacgaa atgttcagca ctacgcatac ttttgacagg aaacgcaacg gatattgagt
     3001 caatatcagg cattctatcg ctcagcttta cagtgacaat gacggctggc gactgaatat
     3061 tagtgcttac agacagcact acatattttc cgtcgatgtt gaaatccttt ctcatatgtc
     3121 accataaata tcaaataatt atagcaatca tttacgcgtt aatggctaat cgccatcttc
     3181 cagcaggcgc accattgccc ctgtttcact atccagggta cggatatagt tcatgacaat
     3241 atttacattg gtccagccac cagcttgcat gatctccggt attgaaactc cagcgcgggc
     3301 catatctcgc gcggctccga cacgggcact gtgtccagac caggccaggt atctctgacc
     3361 agagtcatcc ttagcgccgt aaatcaatcg atgagttgct tcaaaaatcc cttccagggc
     3421 gcgagttgat agctggctgg tggcagatgg cgcggcaaca ccattttttc tgacccggca
     3481 aaacaggtag ttattcggat catcagctac accagagacg gaaatccatc gctcgaccag
     3541 tttagttacc cccaggctaa gtgccttctc tacacctgcg gtgctaacca gcgttttcgt
     3601 tctgccaata tggattaaca ttctcccacc gtcagtacgt gagatatctt taaccctgat
     3661 cctggcaatt tcggctatac gtaacagggt gttataagca atccccagaa atgccagatt
     3721 acgtatatcc tggcagcgat cgctattttc catgagtgaa cgaacctggt cgaaatcagt
     3781 gcgttcgaac gctagagcct gttttgcacg ttcaccggca tcaacgtttt cttttcggat
     3841 ccgccgcata accagtgaaa cagcattgct gtcacttggt cgtggcagcc cggaccgacg
     3901 atgaagcatg tttagctggc ccaaatgttg ctggatagtt tttactgcca gaccgcgcgc
     3961 ctgaagatat agaagataat cgcgaacatc ttcaggttct gcgggaaacc atttccggtt
     4021 attcaacttg caccatgccg cccacgaccg gcaaacggac agaagcattt tccaggtatg
     4081 ctcagaaaac gcctggcgat ccctgaacat gtccatcagg ttcttgcgaa cctcatcact
     4141 cgttgcatcg accggtaatg caggcaaatt ttggtgtacg gtcagtaaat tggacattta
     4201 acactcagat aatggtttta agtaaagtgt acaggatcgg ctctgcccct cgacggtatc
     4261 gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag aatccggggt
     4321 tttttctcct tgacgttaaa gtatagaggt atattaacaa ttttttgttg atacttttat
     4381 tacatttgaa taagaagtaa tacaaaccga aaatgttgaa agtattagtt aaagtggtta
     4441 tgcagttttt gcatttatat atctgttaat agatcaaaaa tcatcgcttc gctgattaat
     4501 taccccagaa ataaggctaa aaaactaatc gcattatcat cctatggttg ttaatttgat
     4561 tcgttcattt gaaggtttgt ggggccaggt tactgccaat ttttcctctt cataaccata
     4621 aaagctagta ttgtagaatc tttattgttc ggagcagtgc ggcgcgaggc acatctgcgt
     4681 ttcaggaacg cgaccggtga agacgaggac gcacggagga gagtcttcct tcggagggct
     4741 gtcacccgct cggcggcttc taatccgtac tagagctcca gcttttgttc cctttagtga
     4801 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4861 ccgctcacaa ttccacacaa cataggagcc ggaagcataa agtgtaaagc ctggggtgcc
     4921 taatgagtga ggtaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4981 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5041 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5101 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5401 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5641 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5941 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6001 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6061 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6961 tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat aattataatt
     7021 taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga aattaaagaa
     7081 aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca acaaatacta
     7141 ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca tcttgtctgt
     7201 gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt aatcgaatgt
     7261 atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac gctttttgtt
     7321 gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt ctaccttctt
     7381 tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag agtaaattcc
     7441 caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc
     7501 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7561 tcgtc
//
