LOCUS       Exported                6809 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Constitutive expression vector for Cre-recombinase containing TRP1 
            selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    pPL5608_TEF1*-Cre_TRP1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6809)
  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 6809)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6809
                     /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"
     promoter        249..529
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             530..1204
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1303..1758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1440..1461)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1652..1671
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1888..1910
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1902..1919
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1903..1919
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1929..1948
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1929..1947
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1966..2213
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2179..2197
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2532..3563)
                     /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(4097..4117)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4097..4115)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4136..4152)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4136..4152)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4149..4171)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4160..4176
                     /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(4184..4214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4367..4384)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4538..5126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4618..4637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5297..6157)
                     /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     5920..5939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6158..6262)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6299..6802
                     /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 gcaccataaa cgacattact atatatataa tataggaagc atttaataga cagcatcgta
      301 atatatgtgt actttgcagt tatgacgcca gatggcagta gtggaagata ttctttattg
      361 aaaaatagct tgtcacctta cgtacaatct tgatccggag cttttctttt tttgccgatt
      421 aagaattaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact
      481 attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat
      541 taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc
      601 cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag
      661 aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa
      721 aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga
      781 tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc
      841 gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa
      901 agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt
      961 tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt
     1021 tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga
     1081 aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt
     1141 ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa
     1201 ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc tatgcggtgt
     1261 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1321 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1381 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1441 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1501 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1561 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1621 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1681 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1741 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     1801 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1861 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     1921 agcgcgcgta atacgactca ctatagggcg aattgggtac cggccgcaaa ttaaagcctt
     1981 cgagcgtccc aaaaccttct caagcaaggt tttcagtata atgttacatg cgtacacgcg
     2041 tctgtacaga aaaaaaagaa aaatttgaaa tataaataac gttcttaata ctaacataac
     2101 tataaaaaaa taaataggga cctagacttc aggttgtcta actccttcct tttcggttag
     2161 agcggatgtg gggggagggc gtgaatgtaa gcgtgacata actaattaca tgactcgagg
     2221 tcgacaattc caaccttacc caagagttcg ccaaactcag acatcacttt angcaaaacc
     2281 gcgccgtgct tcttcctcgg tggcattcat cacgaaatgt tcagcactac gcatactttt
     2341 gacaggaaac gcaacggata ttgagtcaat atcaggcatt ctatcgctca gctttacagt
     2401 gacaatgacg gctggcgact gaatattagt gcttacagac agcactacat attttccgtc
     2461 gatgttgaaa tcctttctca tatgtcacca taaatatcaa ataattatag caatcattta
     2521 cgcgttaatg gctaatcgcc atcttccagc aggcgcacca ttgcccctgt ttcactatcc
     2581 agggtacgga tatagttcat gacaatattt acattggtcc agccaccagc ttgcatgatc
     2641 tccggtattg aaactccagc gcgggccata tctcgcgcgg ctccgacacg ggcactgtgt
     2701 ccagaccagg ccaggtatct ctgaccagag tcatccttag cgccgtaaat caatcgatga
     2761 gttgcttcaa aaatcccttc cagggcgcga gttgatagct ggctggtggc agatggcgcg
     2821 gcaacaccat tttttctgac ccggcaaaac aggtagttat tcggatcatc agctacacca
     2881 gagacggaaa tccatcgctc gaccagttta gttaccccca ggctaagtgc cttctctaca
     2941 cctgcggtgc taaccagcgt tttcgttctg ccaatatgga ttaacattct cccaccgtca
     3001 gtacgtgaga tatctttaac cctgatcctg gcaatttcgg ctatacgtaa cagggtgtta
     3061 taagcaatcc ccagaaatgc cagattacgt atatcctggc agcgatcgct attttccatg
     3121 agtgaacgaa cctggtcgaa atcagtgcgt tcgaacgcta gagcctgttt tgcacgttca
     3181 ccggcatcaa cgttttcttt tcggatccgc cgcataacca gtgaaacagc attgctgtca
     3241 cttggtcgtg gcagcccgga ccgacgatga agcatgttta gctggcccaa atgttgctgg
     3301 atagttttta ctgccagacc gcgcgcctga agatatagaa gataatcgcg aacatcttca
     3361 ggttctgcgg gaaaccattt ccggttattc aacttgcacc atgccgccca cgaccggcaa
     3421 acggacagaa gcattttcca ggtatgctca gaaaacgcct ggcgatccct gaacatgtcc
     3481 atcaggttct tgcgaacctc atcactcgtt gcatcgaccg gtaatgcagg caaattttgg
     3541 tgtacggtca gtaaattgga catttaacac tcagataatg gttttaagta aagtgtacag
     3601 gatcggctct gcccctcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
     3661 gatccactag tttttgtaat taaaacttag attagattgc tatgctttct ttccaatgag
     3721 caagaagtaa aaaaagttgt aatagaacag gaaaaatgaa gctgaaactt gagaaattga
     3781 agaccgtttg ttaactcaaa tatcaatggg aggtcgtcga aagagaacaa aatcgaaaaa
     3841 aaagttttca agagaaagaa acgtgataaa aatttttatt gccttctccg acgaagaaaa
     3901 agggacgagg cggtctcttt ttccttttcc aaacctttag tacgggtaat taacggcacc
     3961 ctagaggaag gaggaggggg aatttagtat gctgtgcttg ggtgttttga agtggtacgg
     4021 cggtgcgcgg agtccgagaa aatctggaag agtaaaaaag gagtagagac attttgaagc
     4081 tatccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     4141 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa
     4201 gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc
     4261 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4321 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4381 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4441 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4501 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4561 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4621 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4681 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4741 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4801 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4861 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4921 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4981 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5041 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5101 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5161 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5221 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5281 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5341 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5401 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5461 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5521 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5581 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5641 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5701 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5761 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5821 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5881 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5941 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6001 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6061 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6121 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6181 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6241 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat
     6301 cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat
     6361 agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac
     6421 tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta
     6481 atgccgaatt gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta
     6541 cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa
     6601 tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct
     6661 tcattccgta actcttctac cttctttatt tactttctaa aatccaaata caaaacataa
     6721 aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg
     6781 tgtaagttac aggcaagcga tccgtccta
//
