LOCUS       Exported                9380 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  S. cerevisae centromic plasmid harboring Fncpf1 under control of 
            TEF1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pUDC175
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9380)
  AUTHORS   Swiat MA, Dashko S, den Ridder M, Wijsman M, van der Oost J, Daran 
            JM Daran-Lapujade P.
  TITLE     FnCpf1, a novel and efficient genome editing tool for Saccharomyces 
            cerevisiae.
  JOURNAL   Nucleic Acids Research (2017), gkx1007
REFERENCE   2  (bases 1 to 9380)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9380
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      1..190
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(220..239)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(221..239)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(249..266)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(249..265)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(258..280)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      406..861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(493..512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     703..724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(964..1638)
                     /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"
     promoter        complement(1639..1919)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(1940..1959)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2059..2081
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2119..2137)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2178..2681
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2718..2822
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2823..3683
                     /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     complement(3041..3060)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3854..4442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4343..4362
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4596..4613
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4730..4751
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4766..4796
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4804..4820
                     /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)."
     primer_bind     4809..4831
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4828..4844
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4828..4844
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4863..4883
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4865..4883
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4901..5299
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             9204..9251
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9258..9338
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(9351..9369)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
       61 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
      121 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
      181 acgctcgaag gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac
      241 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      301 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      361 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
      421 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      481 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      541 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      601 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
      661 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      721 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
      781 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      841 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
      901 ttcacaccgc atagatcggc aagtgcacaa acaatactta aataaatact actcagtaat
      961 aacctatttc ttagcatttt tgacgaaatt tgctattttg ttagagtctt ttacaccatt
     1021 tgtctccaca cctccgctta catcaacacc aataacgcca tttaatctaa gcgcatcacc
     1081 aacattttct ggcgtcagtc caccagctaa cataaaatgt aagctttcgg ggctctcttg
     1141 ccttccaacc cagtcagaaa tcgagttcca atccaaaagt tcacctgtcc cacctgcttc
     1201 tgaatcaaac aagggaataa acgaatgagg tttctgtgaa gctgcactga gtagtatgtt
     1261 gcagtctttt ggaaatacga gtcttttaat aactggcaaa ccgaggaact cttggtattc
     1321 ttgccacgac tcatctccat gcagttggac gatatcaatg ccgtaatcat tgaccagagc
     1381 caaaacatcc tccttaggtt gattacgaaa cacgccaacc aagtatttcg gagtgcctga
     1441 actattttta tatgctttta caagacttga aattttcctt gcaataaccg ggtcaattgt
     1501 tctctttcta ttgggcacac atataatacc cagcaagtca gcatcggaat ctagagcaca
     1561 ttctgcggcc tctgtgctct gcaagccgca aactttcacc aatggaccag aactacctgt
     1621 gaaattaata acagacatac tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt
     1681 gctcaatagt caccaatgcc ctccctcttg gccctctcct tttctttttt cgaccgaatt
     1741 aattcttaat cggcaaaaaa agaaaagctc cggatcaaga ttgtacgtaa ggtgacaagc
     1801 tatttttcaa taaagaatat cttccactac tgccatctgg cgtcataact gcaaagtaca
     1861 catatattac gatgctgttc tattaaatgc ttcctatatt atatatatag taatgtcgtg
     1921 atctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     1981 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2041 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2101 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     2161 taatggtttc ttagacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg
     2221 atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt
     2281 tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg
     2341 tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag
     2401 attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg
     2461 tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga
     2521 gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa
     2581 caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt atatatttat
     2641 attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca
     2701 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2761 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2821 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2881 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2941 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3001 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3061 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3121 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3181 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3241 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3301 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3361 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3421 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3481 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3541 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3601 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3661 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3721 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3781 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3841 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3901 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3961 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4021 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4081 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4141 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4201 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4261 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4321 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4381 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4441 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4501 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4561 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4621 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4681 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     4741 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4801 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     4861 gcgcaattaa ccctcactaa agggaacaaa agctggagct catagcttca aaatgtttct
     4921 actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa
     4981 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
     5041 cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa
     5101 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt gatttttttc
     5161 tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt
     5221 cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt agaaagaaag
     5281 catagcaatc taatctaagt tttatgagca tctaccagga gttcgtcaac aagtattcac
     5341 tgagtaagac actgcggttc gagctgatcc cacagggcaa gacactggag aacatcaagg
     5401 cccgaggcct gattctggac gatgagaagc gggcaaaaga ctataagaaa gccaagcaga
     5461 tcattgataa ataccaccag ttctttatcg aggaaattct gagctccgtg tgcatcagtg
     5521 aggatctgct gcagaattac tcagacgtgt acttcaagct gaagaagagc gacgatgaca
     5581 acctgcagaa ggacttcaag tccgccaagg acaccatcaa gaaacagatt agcgagtaca
     5641 tcaaggactc cgaaaagttt aaaaatctgt tcaaccagaa tctgatcgat gctaagaaag
     5701 gccaggagtc cgacctgatc ctgtggctga aacagtctaa ggacaatggg attgaactgt
     5761 tcaaggctaa ctccgatatc actgatattg acgaggcact ggaaatcatc aagagcttca
     5821 agggatggac cacatacttt aaaggcttcc acgagaaccg caagaacgtg tactccagca
     5881 acgacattcc tacctccatc atctaccgaa tcgtcgatga caatctgcca aagttcctgg
     5941 agaacaaggc caaatatgaa tctctgaagg acaaagctcc cgaggcaatt aattacgaac
     6001 agatcaagaa agatctggct gaggaactga cattcgatat cgactataag actagcgagg
     6061 tgaaccagag ggtcttttcc ctggacgagg tgtttgaaat cgccaatttc aacaattacc
     6121 tgaaccagtc cggcattact aaattcaata ccatcattgg cgggaagttt gtgaacgggg
     6181 agaataccaa gcgcaaggga attaacgaat acatcaatct gtatagccag cagatcaacg
     6241 acaaaactct gaagaaatac aagatgtctg tgctgttcaa acagatcctg agtgataccg
     6301 agtccaagtc ttttgtcatt gataaactgg aagatgactc agacgtggtc actaccatgc
     6361 agagctttta tgagcagatc gccgctttca agacagtgga ggaaaaatct attaaggaaa
     6421 ctctgagtct gctgttcgat gacctgaaag cccagaagct ggacctgagt aagatctact
     6481 tcaaaaacga taagagtctg acagacctgt cacagcaggt gtttgatgac tattccgtga
     6541 ttgggaccgc cgtcctggag tacattacac agcagatcgc tccaaagaac ctggataatc
     6601 cctctaagaa agagcaggaa ctgatcgcta agaaaaccga gaaggcaaaa tatctgagtc
     6661 tggaaacaat taagctggca ctggaggagt tcaacaagca cagggatatt gacaaacagt
     6721 gccgctttga ggaaatcctg gccaacttcg cagccatccc catgattttt gatgagatcg
     6781 cccagaacaa agacaatctg gctcagatca gtattaagta ccagaaccag ggcaagaaag
     6841 acctgctgca ggcttcagca gaagatgacg tgaaagccat caaggatctg ctggaccaga
     6901 ccaacaatct gctgcacaag ctgaaaatct tccatattag tcagtcagag gataaggcta
     6961 atatcctgga taaagacgaa cacttctacc tggtgttcga ggaatgttac ttcgagctgg
     7021 caaacattgt ccccctgtat aacaagatta ggaactacat cacacagaag ccttactctg
     7081 acgagaagtt taaactgaac ttcgaaaata gtaccctggc caacgggtgg gataagaaca
     7141 aggagcctga caacacagct atcctgttca tcaaggatga caagtactat ctgggagtga
     7201 tgaataagaa aaacaataag atcttcgatg acaaagccat taaggagaac aaaggggaag
     7261 gatacaagaa aatcgtgtat aagctgctgc ccggcgcaaa taagatgctg cctaaggtgt
     7321 tcttcagcgc caagagtatc aaattctaca acccatccga ggacatcctg cggattagaa
     7381 atcactcaac acatactaag aacgggagcc cccagaaggg atatgagaaa tttgagttca
     7441 acatcgagga ttgcaggaag tttattgact tctacaagca gagcatctcc aaacaccctg
     7501 aatggaagga ttttggcttc cggttttccg acacacagag atataactct atcgacgagt
     7561 tctaccgcga ggtggaaaat caggggtata agctgacttt tgagaacatt tctgaaagtt
     7621 acatcgacag cgtggtcaat cagggaaagc tgtacctgtt ccagatctat aacaaagatt
     7681 tttcagcata cagcaagggc agaccaaacc tgcatacact gtactggaag gccctgttcg
     7741 atgagaggaa tctgcaggac gtggtctata aactgaacgg agaggccgaa ctgttttacc
     7801 ggaagcagtc tattcctaag aaaatcactc acccagctaa ggaggccatc gctaacaaga
     7861 acaaggacaa tcctaagaaa gagagcgtgt tcgaatacga tctgattaag gacaagcggt
     7921 tcaccgaaga taagttcttt ttccattgtc caatcaccat taacttcaag tcaagcggcg
     7981 ctaacaagtt caacgacgag atcaatctgc tgctgaagga aaaagcaaac gatgtgcaca
     8041 tcctgagcat tgaccgagga gagcggcatc tggcctacta taccctggtg gatggcaaag
     8101 ggaatatcat taagcaggat acattcaaca tcattggcaa tgaccggatg aaaaccaact
     8161 accacgataa actggctgca atcgagaagg atagagactc agctaggaag gactggaaga
     8221 aaatcaacaa cattaaggag atgaaggaag gctatctgag ccaggtggtc catgagattg
     8281 caaagctggt catcgaatac aatgccattg tggtgttcga ggatctgaac ttcggcttta
     8341 agagggggcg ctttaaggtg gaaaaacagg tctatcagaa gctggagaaa atgctgatcg
     8401 aaaagctgaa ttacctggtg tttaaagata acgagttcga caagaccgga ggcgtcctga
     8461 gagcctacca gctgacagct ccctttgaaa ctttcaagaa aatgggaaaa cagacaggca
     8521 tcatctacta tgtgccagcc ggattcactt ccaagatctg ccccgtgacc ggctttgtca
     8581 accagctgta ccctaaatat gagtcagtga gcaagtccca ggaatttttc agcaagttcg
     8641 ataagatctg ttataatctg gacaaggggt acttcgagtt ttccttcgat tacaagaact
     8701 tcggcgacaa ggccgctaag gggaaatgga ccattgcctc cttcggatct cgcctgatca
     8761 actttcgaaa ttccgataaa aaccacaatt gggacactag ggaggtgtac ccaaccaagg
     8821 agctggaaaa gctgctgaaa gactactcta tcgagtatgg acatggcgaa tgcatcaagg
     8881 cagccatctg tggcgagagt gataagaaat ttttcgccaa gctgacctca gtgctgaata
     8941 caatcctgca gatgcggaac tcaaagaccg ggacagaact ggactatctg attagccccg
     9001 tggctgatgt caacggaaac ttcttcgaca gcagacaggc acccaaaaat atgcctcagg
     9061 atgcagacgc caacggggcc taccacatcg ggctgaaggg actgatgctg ctgggccgga
     9121 tcaagaacaa tcaggagggg aagaagctga acctggtcat taagaacgag gaatacttcg
     9181 agtttgtcca gaatagaaat aacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa
     9241 aaaagaaaaa gggatcctac ccatacgatg ttccagatta cgcttatccc tacgacgtgc
     9301 ctgattatgc atacccatat gatgtccccg actatgccta aattagttat gtcacgctta
     9361 cattcacgcc ctccccccac
//
