LOCUS       Exported                7772 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Constitutive expression vector for Cre-recombinase containing MET15 
            selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    pPL5628_TEF1*-Cre_MET15
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7772)
  AUTHORS   MacDonald C, Piper RC
  TITLE     Puromycin and Methotrexate Resistance Cassettes and Optimized 
            cre-recombinase Expression Plasmids for use in Yeast.
  JOURNAL   Yeast. 2015 Feb 16. doi: 10.1002/yea.3069.
  PUBMED    25688547
REFERENCE   2  (bases 1 to 7772)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7772
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     35..53
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(91..113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     213..232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             588..1922
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     rep_origin      complement(2342..2722)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2404..2425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2616..2635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2852..2874
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2866..2883
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2867..2883
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2893..2912
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2893..2911
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2930..3177
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3143..3161
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(3496..4527)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(5061..5081)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5061..5079)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5100..5116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5100..5116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5113..5135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5124..5140
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(5148..5178)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5193..5214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5331..5348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5502..6090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5582..5601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6261..7121)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6884..6903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7122..7226)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7263..7766
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
       61 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
      121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
      181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
      241 gcaccacgct tttcatcgag gtcacatgat cgcaaaatgg caaatggcac gtgaagctgt
      301 cgatattggg gaactgtggt ggttggcaaa tgactaatta agttagtcaa ggcgccatcc
      361 tcatgaaaac tgtgtaacat aataaccgaa gtgtcgaaaa ggtggcacct tgtccaattg
      421 aacacgctcg atgaaaaaaa taagatatat ataaggttaa gtaaagcgtc tgttagaaag
      481 gaagtttttc ctttttcttg ctctcttgtc ttttcatcta ctatttcctt cgtgtaatac
      541 agggtcgtca gatacataga tacaattcta ttacccccat ccatacaatg ccatctcatt
      601 tcgatactgt tcaactacac gccggccaag agaaccctgg tgacaatgct cacagatcca
      661 gagctgtacc aatttacgcc accacttctt atgttttcga aaactctaag catggttcgc
      721 aattgtttgg tctagaagtt ccaggttacg tctattcccg tttccaaaac ccaaccagta
      781 atgttttgga agaaagaatt gctgctttag aaggtggtgc tgctgctttg gctgtttcct
      841 ccggtcaagc cgctcaaacc cttgccatcc aaggtttggc acacactggt gacaacatcg
      901 tttccacttc ttacttatac ggtggtactt ataaccagtt caaaatctcg ttcaaaagat
      961 ttggtatcga ggctagattt gttgaaggtg acaatccaga agaattcgaa aaggtctttg
     1021 atgaaagaac caaggctgtt tatttggaaa ccattggtaa tccaaagtac aatgttccgg
     1081 attttgaaaa aattgttgca attgctcaca aacacggtat tccagttgtc gttgacaaca
     1141 catttggtgc cggtggttac ttctgtcagc caattaaata cggtgctgat attgtaacac
     1201 attctgctac caaatggatt ggtggtcatg gtactactat cggtggtatt attgttgact
     1261 ctggtaagtt cccatggaag gactacccag aaaagttccc tcaattctct caacctgccg
     1321 aaggatatca cggtactatc tacaatgaag cctacggtaa cttggcatac atcgttcatg
     1381 ttagaactga actattaaga gatttgggtc cattgatgaa cccatttgcc tctttcttgc
     1441 tactacaagg tgttgaaaca ttatctttga gagctgaaag acacggtgaa aatgcattga
     1501 agttagccaa atggttagaa caatccccat acgtatcttg ggtttcatac cctggtttag
     1561 catctcattc tcatcatgaa aatgctaaga agtatctatc taacggtttc ggtggtgtct
     1621 tatctttcgg tgtaaaagac ttaccaaatg ccgacaagga aactgaccca ttcaaacttt
     1681 ctggtgctca agttgttgac aatttaaagc ttgcctctaa cttggccaat gttggtgatg
     1741 ccaagacctt agtcattgct ccatacttca ctacccacaa acaattaaat gacaaagaaa
     1801 agttggcatc tggtgttacc aaggacttaa ttcgtgtctc tgttggtatc gaatttattg
     1861 atgacattat tgcagacttc cagcaatctt ttgaaactgt tttcgctggc caaaaaccat
     1921 gagtgtgcgt aatgagttgt aaaattatgt ataaacctac tttctctcac aagtactata
     1981 cttttataaa acgaacttta ttgaaatgaa tatccttttt ttcccttgtt acatgtcgtg
     2041 actcgtactt tgaacctaaa ttgttctaac atcaaagaac agtgttaatt cgcagtcgag
     2101 aagaaaaata tggtgaacaa gactcatcta cttcatgaga ctactttacg cctcctataa
     2161 agctgtcaca ctggataaat ttattgtagg accaagttac aaaagaggat gatggaggtt
     2221 tctttacaat aaagaagcac atgtgtgtta acgtttttag tatttgcttg ttatgtaaat
     2281 caggaaaact tcgcgggatt tggttggatg cgttaaatca gctcattttt taaccaatag
     2341 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2401 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2461 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2521 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2581 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2641 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2701 aatgcgccgc tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa
     2761 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     2821 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     2881 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccggccg caaattaaag
     2941 ccttcgagcg tcccaaaacc ttctcaagca aggttttcag tataatgtta catgcgtaca
     3001 cgcgtctgta cagaaaaaaa agaaaaattt gaaatataaa taacgttctt aatactaaca
     3061 taactataaa aaaataaata gggacctaga cttcaggttg tctaactcct tccttttcgg
     3121 ttagagcgga tgtgggggga gggcgtgaat gtaagcgtga cataactaat tacatgactc
     3181 gaggtcgaca attccaacct tacccaagag ttcgccaaac tcagacatca ctttangcaa
     3241 aaccgcgccg tgcttcttcc tcggtggcat tcatcacgaa atgttcagca ctacgcatac
     3301 ttttgacagg aaacgcaacg gatattgagt caatatcagg cattctatcg ctcagcttta
     3361 cagtgacaat gacggctggc gactgaatat tagtgcttac agacagcact acatattttc
     3421 cgtcgatgtt gaaatccttt ctcatatgtc accataaata tcaaataatt atagcaatca
     3481 tttacgcgtt aatggctaat cgccatcttc cagcaggcgc accattgccc ctgtttcact
     3541 atccagggta cggatatagt tcatgacaat atttacattg gtccagccac cagcttgcat
     3601 gatctccggt attgaaactc cagcgcgggc catatctcgc gcggctccga cacgggcact
     3661 gtgtccagac caggccaggt atctctgacc agagtcatcc ttagcgccgt aaatcaatcg
     3721 atgagttgct tcaaaaatcc cttccagggc gcgagttgat agctggctgg tggcagatgg
     3781 cgcggcaaca ccattttttc tgacccggca aaacaggtag ttattcggat catcagctac
     3841 accagagacg gaaatccatc gctcgaccag tttagttacc cccaggctaa gtgccttctc
     3901 tacacctgcg gtgctaacca gcgttttcgt tctgccaata tggattaaca ttctcccacc
     3961 gtcagtacgt gagatatctt taaccctgat cctggcaatt tcggctatac gtaacagggt
     4021 gttataagca atccccagaa atgccagatt acgtatatcc tggcagcgat cgctattttc
     4081 catgagtgaa cgaacctggt cgaaatcagt gcgttcgaac gctagagcct gttttgcacg
     4141 ttcaccggca tcaacgtttt cttttcggat ccgccgcata accagtgaaa cagcattgct
     4201 gtcacttggt cgtggcagcc cggaccgacg atgaagcatg tttagctggc ccaaatgttg
     4261 ctggatagtt tttactgcca gaccgcgcgc ctgaagatat agaagataat cgcgaacatc
     4321 ttcaggttct gcgggaaacc atttccggtt attcaacttg caccatgccg cccacgaccg
     4381 gcaaacggac agaagcattt tccaggtatg ctcagaaaac gcctggcgat ccctgaacat
     4441 gtccatcagg ttcttgcgaa cctcatcact cgttgcatcg accggtaatg caggcaaatt
     4501 ttggtgtacg gtcagtaaat tggacattta acactcagat aatggtttta agtaaagtgt
     4561 acaggatcgg ctctgcccct cgacggtatc gataagcttg atatcgaatt cctgcagccc
     4621 gggggatcca ctagtttttg taattaaaac ttagattaga ttgctatgct ttctttccaa
     4681 tgagcaagaa gtaaaaaaag ttgtaataga acaggaaaaa tgaagctgaa acttgagaaa
     4741 ttgaagaccg tttgttaact caaatatcaa tgggaggtcg tcgaaagaga acaaaatcga
     4801 aaaaaaagtt ttcaagagaa agaaacgtga taaaaatttt tattgccttc tccgacgaag
     4861 aaaaagggac gaggcggtct ctttttcctt ttccaaacct ttagtacggg taattaacgg
     4921 caccctagag gaaggaggag ggggaattta gtatgctgtg cttgggtgtt ttgaagtggt
     4981 acggcggtgc gcggagtccg agaaaatctg gaagagtaaa aaaggagtag agacattttg
     5041 aagctatcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     5101 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5161 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5221 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5281 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5341 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5401 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5461 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5521 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5581 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5641 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5701 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5761 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5821 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5881 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5941 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6001 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6061 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6121 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6181 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6241 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6301 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6361 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6421 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6481 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6541 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6601 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6661 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6721 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6781 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6841 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6901 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6961 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7021 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7081 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7141 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7201 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtccttt
     7261 tcatcacgtg ctataaaaat aattataatt taaatttttt aatataaata tataaattaa
     7321 aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg aagatgtaaa
     7381 agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct gctctcaggt
     7441 attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa atcctgtgat
     7501 tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt tcttgtctaa
     7561 taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt ttattttttt
     7621 ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca aatacaaaac
     7681 ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa agatacgagg
     7741 cgcgtgtaag ttacaggcaa gcgatccgtc ta
//
