LOCUS       Exported                7868 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Constitutive expression vector for Cre-recombinase containing ADE2 
            selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    pPL5606_TEF1*-Cre_ADE2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7868)
  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 7868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7868
                     /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             480..2195
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(2362..2817)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2499..2520)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2711..2730
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2947..2969
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2961..2978
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2962..2978
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2988..3007
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2988..3006
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3025..3272
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3238..3256
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(3591..4622)
                     /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(5156..5176)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5156..5174)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5195..5211)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5195..5211)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5208..5230)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5219..5235
                     /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(5243..5273)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5426..5443)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5597..6185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5677..5696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6356..7216)
                     /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     6979..6998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7217..7321)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7358..7861
                     /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 gcaccacgcc gtcatgattg attattacag ctatgctgac aaatgactct tgttgcatgg
      301 ctacgaaccg ggtaatacta agtgattgac tcttgctgac cttttattaa gaactaaatg
      361 gacaatatta tggagcattt catgtataaa ttggtgcgta aaatcgttgg atctctcttc
      421 taagtacatc ctactataac aatcaagaaa aacaagaaaa tcggacaaaa caatcaagta
      481 tggattctag aacagttggt atattaggag ggggacaatt gggacgtatg attgttgagg
      541 cagcaaacag gctcaacatt aagacggtaa tactagatgc tgaaaattct cctgccaaac
      601 aaataagcaa ctccaatgac cacgttaatg gctccttttc caatcctctt gatatcgaaa
      661 aactagctga aaaatgtgat gtgctaacga ttgagattga gcatgttgat gttcctacac
      721 taaagaatct tcaagtaaaa catcccaaat taaaaattta cccttctcca gaaacaatca
      781 gattgataca agacaaatat attcaaaaag agcatttaat caaaaatggt atagcagtta
      841 cccaaagtgt tcctgtggaa caagccagtg agacgtccct attgaatgtt ggaagagatt
      901 tgggttttcc attcgtcttg aagtcgagga ctttggcata cgatggaaga ggtaacttcg
      961 ttgtaaagaa taaggaaatg attccggaag ctttggaagt actgaaggat cgtcctttgt
     1021 acgccgaaaa atgggcacca tttactaaag aattagcagt catgattgtg agatctgtta
     1081 acggtttagt gttttcttac ccaattgtag agactatcca caaggacaat atttgtgact
     1141 tatgttatgc gcctgctaga gttccggact ccgttcaact taaggcgaag ttgttggcag
     1201 aaaatgcaat caaatctttt cccggttgtg gtatatttgg tgtggaaatg ttctatttag
     1261 aaacagggga attgcttatt aacgaaattg ccccaaggcc tcacaactct ggacattata
     1321 ccattgatgc ttgcgtcact tctcaatttg aagctcattt gagatcaata ttggatttgc
     1381 caatgccaaa gaatttcaca tctttctcca ccattacaac gaacgccatt atgctaaatg
     1441 ttcttggaga caaacataca aaagataaag agctagaaac ttgcgaaaga gcattggcga
     1501 ctccaggttc ctcagtgtac ttatatggaa aagagtctag acctaacaga aaagtaggtc
     1561 acataaatat tattgcctcc agtatggcgg aatgtgaaca aaggctgaac tacattacag
     1621 gtagaactga tattccaatc aaaatctctg tcgctcaaaa gttggacttg gaagcaatgg
     1681 tcaaaccatt ggttggaatc atcatgggat cagactctga cttgccggta atgtctgccg
     1741 catgtgcggt tttaaaagat tttggcgttc catttgaagt gacaatagtc tctgctcata
     1801 gaactccaca taggatgtca gcatatgcta tttccgcaag caagcgtgga attaaaacaa
     1861 ttatcgctgg agctggtggg gctgctcact tgccaggtat ggtggctgca atgacaccac
     1921 ttcctgtcat cggtgtgccc gtaaaaggtt cttgtctaga tggagtagat tctttacatt
     1981 caattgtgca aatgcctaga ggtgttccag tagctaccgt cgctattaat aatagtacga
     2041 acgctgcgct gttggctgtc agactgcttg gcgcttatga ttcaagttat acaacgaaaa
     2101 tggaacagtt tttattaaag caagaagaag aagttcttgt caaagcacaa aagttagaaa
     2161 ctgtcggtta cgaagcttat ctagaaaaca agtaatatat aagtttattg atatacttgt
     2221 acagcaaata attataaaat gatataccta ttttttaggc tttgttatga ttacatcaaa
     2281 tgtggacttc atacatagaa atcaacgctt acacggtgtg aaataccgca cagatgcgta
     2341 aggagaaaat accgcatcag gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa
     2401 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2461 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2521 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2581 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2641 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2701 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2761 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcgc
     2821 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     2881 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     2941 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     3001 tatagggcga attgggtacc ggccgcaaat taaagccttc gagcgtccca aaaccttctc
     3061 aagcaaggtt ttcagtataa tgttacatgc gtacacgcgt ctgtacagaa aaaaaagaaa
     3121 aatttgaaat ataaataacg ttcttaatac taacataact ataaaaaaat aaatagggac
     3181 ctagacttca ggttgtctaa ctccttcctt ttcggttaga gcggatgtgg ggggagggcg
     3241 tgaatgtaag cgtgacataa ctaattacat gactcgaggt cgacaattcc aaccttaccc
     3301 aagagttcgc caaactcaga catcacttta ngcaaaaccg cgccgtgctt cttcctcggt
     3361 ggcattcatc acgaaatgtt cagcactacg catacttttg acaggaaacg caacggatat
     3421 tgagtcaata tcaggcattc tatcgctcag ctttacagtg acaatgacgg ctggcgactg
     3481 aatattagtg cttacagaca gcactacata ttttccgtcg atgttgaaat cctttctcat
     3541 atgtcaccat aaatatcaaa taattatagc aatcatttac gcgttaatgg ctaatcgcca
     3601 tcttccagca ggcgcaccat tgcccctgtt tcactatcca gggtacggat atagttcatg
     3661 acaatattta cattggtcca gccaccagct tgcatgatct ccggtattga aactccagcg
     3721 cgggccatat ctcgcgcggc tccgacacgg gcactgtgtc cagaccaggc caggtatctc
     3781 tgaccagagt catccttagc gccgtaaatc aatcgatgag ttgcttcaaa aatcccttcc
     3841 agggcgcgag ttgatagctg gctggtggca gatggcgcgg caacaccatt ttttctgacc
     3901 cggcaaaaca ggtagttatt cggatcatca gctacaccag agacggaaat ccatcgctcg
     3961 accagtttag ttacccccag gctaagtgcc ttctctacac ctgcggtgct aaccagcgtt
     4021 ttcgttctgc caatatggat taacattctc ccaccgtcag tacgtgagat atctttaacc
     4081 ctgatcctgg caatttcggc tatacgtaac agggtgttat aagcaatccc cagaaatgcc
     4141 agattacgta tatcctggca gcgatcgcta ttttccatga gtgaacgaac ctggtcgaaa
     4201 tcagtgcgtt cgaacgctag agcctgtttt gcacgttcac cggcatcaac gttttctttt
     4261 cggatccgcc gcataaccag tgaaacagca ttgctgtcac ttggtcgtgg cagcccggac
     4321 cgacgatgaa gcatgtttag ctggcccaaa tgttgctgga tagtttttac tgccagaccg
     4381 cgcgcctgaa gatatagaag ataatcgcga acatcttcag gttctgcggg aaaccatttc
     4441 cggttattca acttgcacca tgccgcccac gaccggcaaa cggacagaag cattttccag
     4501 gtatgctcag aaaacgcctg gcgatccctg aacatgtcca tcaggttctt gcgaacctca
     4561 tcactcgttg catcgaccgg taatgcaggc aaattttggt gtacggtcag taaattggac
     4621 atttaacact cagataatgg ttttaagtaa agtgtacagg atcggctctg cccctcgacg
     4681 gtatcgataa gcttgatatc gaattcctgc agcccggggg atccactagt ttttgtaatt
     4741 aaaacttaga ttagattgct atgctttctt tccaatgagc aagaagtaaa aaaagttgta
     4801 atagaacagg aaaaatgaag ctgaaacttg agaaattgaa gaccgtttgt taactcaaat
     4861 atcaatggga ggtcgtcgaa agagaacaaa atcgaaaaaa aagttttcaa gagaaagaaa
     4921 cgtgataaaa atttttattg ccttctccga cgaagaaaaa gggacgaggc ggtctctttt
     4981 tccttttcca aacctttagt acgggtaatt aacggcaccc tagaggaagg aggaggggga
     5041 atttagtatg ctgtgcttgg gtgttttgaa gtggtacggc ggtgcgcgga gtccgagaaa
     5101 atctggaaga gtaaaaaagg agtagagaca ttttgaagct atccagcttt tgttcccttt
     5161 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5221 gttatccgct cacaattcca cacaacatag gagccggaag cataaagtgt aaagcctggg
     5281 gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5341 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5401 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5461 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5521 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5581 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5641 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5701 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5761 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5821 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5881 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5941 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6001 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6061 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6121 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6181 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6241 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6301 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6361 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6421 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6481 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6541 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6601 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6661 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6721 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6781 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6841 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6901 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6961 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7021 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7081 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7141 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7201 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7261 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7321 gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc acgtgctata aaaataatta
     7381 taatttaaat tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta
     7441 aagaaaaaat agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa
     7501 tactaccttt tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg
     7561 tctgtgtaga agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg
     7621 aatgtatatc tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt
     7681 ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc
     7741 ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa
     7801 attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat
     7861 ccgtccta
//
