LOCUS       V012542                 9446 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V012542
VERSION     V012542
KEYWORDS    pY004 (pcDNA3.1-hFnCpf1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9446)
  AUTHORS   Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS,
            Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin
            EV, Zhang F
  TITLE     Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas
            System.
  JOURNAL   Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi:
            10.1016/j.cell.2015.09.038.
   PUBMED   26422227
REFERENCE   2  (bases 1 to 9446)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9446)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2015
            Sep 23. pii: S0092-8674(15)01200-3. doi:
            10.1016/j.cell.2015.09.038."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        235..614
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             929..4828
                     /gene="cas12a"
                     /label="CRISPR-associated endonuclease Cas12a"
                     /note="CRISPR-associated endonuclease Cas12a from
                     Francisella tularensis subsp. novicida (strain U112).
                     Accession#: A0Q7Q2"
     CDS             4829..4876
                     /label="nucleoplasmin NLS"
                     /note="bipartite nuclear localization signal from
                     nucleoplasmin"
     CDS             4883..4963
                     /label="3xHA"
                     /note="three tandem HA epitope tags"
     polyA_signal    5043..5267
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5313..5741
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5755..6084
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             6151..6942
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    7119..7252
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7289..7305)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7313..7329)
                     /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(7337..7367)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7382..7403)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(7520..7537)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7691..8279)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8453..9310)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9311..9415)
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgccaccat gagcatctac caggagttcg tcaacaagta
      961 ttcactgagt aagacactgc ggttcgagct gatcccacag ggcaagacac tggagaacat
     1021 caaggcccga ggcctgattc tggacgatga gaagcgggca aaagactata agaaagccaa
     1081 gcagatcatt gataaatacc accagttctt tatcgaggaa attctgagct ccgtgtgcat
     1141 cagtgaggat ctgctgcaga attactcaga cgtgtacttc aagctgaaga agagcgacga
     1201 tgacaacctg cagaaggact tcaagtccgc caaggacacc atcaagaaac agattagcga
     1261 gtacatcaag gactccgaaa agtttaaaaa tctgttcaac cagaatctga tcgatgctaa
     1321 gaaaggccag gagtccgacc tgatcctgtg gctgaaacag tctaaggaca atgggattga
     1381 actgttcaag gctaactccg atatcactga tattgacgag gcactggaaa tcatcaagag
     1441 cttcaaggga tggaccacat actttaaagg cttccacgag aaccgcaaga acgtgtactc
     1501 cagcaacgac attcctacct ccatcatcta ccgaatcgtc gatgacaatc tgccaaagtt
     1561 cctggagaac aaggccaaat atgaatctct gaaggacaaa gctcccgagg caattaatta
     1621 cgaacagatc aagaaagatc tggctgagga actgacattc gatatcgact ataagactag
     1681 cgaggtgaac cagagggtct tttccctgga cgaggtgttt gaaatcgcca atttcaacaa
     1741 ttacctgaac cagtccggca ttactaaatt caataccatc attggcggga agtttgtgaa
     1801 cggggagaat accaagcgca agggaattaa cgaatacatc aatctgtata gccagcagat
     1861 caacgacaaa actctgaaga aatacaagat gtctgtgctg ttcaaacaga tcctgagtga
     1921 taccgagtcc aagtcttttg tcattgataa actggaagat gactcagacg tggtcactac
     1981 catgcagagc ttttatgagc agatcgccgc tttcaagaca gtggaggaaa aatctattaa
     2041 ggaaactctg agtctgctgt tcgatgacct gaaagcccag aagctggacc tgagtaagat
     2101 ctacttcaaa aacgataaga gtctgacaga cctgtcacag caggtgtttg atgactattc
     2161 cgtgattggg accgccgtcc tggagtacat tacacagcag atcgctccaa agaacctgga
     2221 taatccctct aagaaagagc aggaactgat cgctaagaaa accgagaagg caaaatatct
     2281 gagtctggaa acaattaagc tggcactgga ggagttcaac aagcacaggg atattgacaa
     2341 acagtgccgc tttgaggaaa tcctggccaa cttcgcagcc atccccatga tttttgatga
     2401 gatcgcccag aacaaagaca atctggctca gatcagtatt aagtaccaga accagggcaa
     2461 gaaagacctg ctgcaggctt cagcagaaga tgacgtgaaa gccatcaagg atctgctgga
     2521 ccagaccaac aatctgctgc acaagctgaa aatcttccat attagtcagt cagaggataa
     2581 ggctaatatc ctggataaag acgaacactt ctacctggtg ttcgaggaat gttacttcga
     2641 gctggcaaac attgtccccc tgtataacaa gattaggaac tacatcacac agaagcctta
     2701 ctctgacgag aagtttaaac tgaacttcga aaatagtacc ctggccaacg ggtgggataa
     2761 gaacaaggag cctgacaaca cagctatcct gttcatcaag gatgacaagt actatctggg
     2821 agtgatgaat aagaaaaaca ataagatctt cgatgacaaa gccattaagg agaacaaagg
     2881 ggaaggatac aagaaaatcg tgtataagct gctgcccggc gcaaataaga tgctgcctaa
     2941 ggtgttcttc agcgccaaga gtatcaaatt ctacaaccca tccgaggaca tcctgcggat
     3001 tagaaatcac tcaacacata ctaagaacgg gagcccccag aagggatatg agaaatttga
     3061 gttcaacatc gaggattgca ggaagtttat tgacttctac aagcagagca tctccaaaca
     3121 ccctgaatgg aaggattttg gcttccggtt ttccgacaca cagagatata actctatcga
     3181 cgagttctac cgcgaggtgg aaaatcaggg gtataagctg acttttgaga acatttctga
     3241 aagttacatc gacagcgtgg tcaatcaggg aaagctgtac ctgttccaga tctataacaa
     3301 agatttttca gcatacagca agggcagacc aaacctgcat acactgtact ggaaggccct
     3361 gttcgatgag aggaatctgc aggacgtggt ctataaactg aacggagagg ccgaactgtt
     3421 ttaccggaag cagtctattc ctaagaaaat cactcaccca gctaaggagg ccatcgctaa
     3481 caagaacaag gacaatccta agaaagagag cgtgttcgaa tacgatctga ttaaggacaa
     3541 gcggttcacc gaagataagt tctttttcca ttgtccaatc accattaact tcaagtcaag
     3601 cggcgctaac aagttcaacg acgagatcaa tctgctgctg aaggaaaaag caaacgatgt
     3661 gcacatcctg agcattgacc gaggagagcg gcatctggcc tactataccc tggtggatgg
     3721 caaagggaat atcattaagc aggatacatt caacatcatt ggcaatgacc ggatgaaaac
     3781 caactaccac gataaactgg ctgcaatcga gaaggataga gactcagcta ggaaggactg
     3841 gaagaaaatc aacaacatta aggagatgaa ggaaggctat ctgagccagg tggtccatga
     3901 gattgcaaag ctggtcatcg aatacaatgc cattgtggtg ttcgaggatc tgaacttcgg
     3961 ctttaagagg gggcgcttta aggtggaaaa acaggtctat cagaagctgg agaaaatgct
     4021 gatcgaaaag ctgaattacc tggtgtttaa agataacgag ttcgacaaga ccggaggcgt
     4081 cctgagagcc taccagctga cagctccctt tgaaactttc aagaaaatgg gaaaacagac
     4141 aggcatcatc tactatgtgc cagccggatt cacttccaag atctgccccg tgaccggctt
     4201 tgtcaaccag ctgtacccta aatatgagtc agtgagcaag tcccaggaat ttttcagcaa
     4261 gttcgataag atctgttata atctggacaa ggggtacttc gagttttcct tcgattacaa
     4321 gaacttcggc gacaaggccg ctaaggggaa atggaccatt gcctccttcg gatctcgcct
     4381 gatcaacttt cgaaattccg ataaaaacca caattgggac actagggagg tgtacccaac
     4441 caaggagctg gaaaagctgc tgaaagacta ctctatcgag tatggacatg gcgaatgcat
     4501 caaggcagcc atctgtggcg agagtgataa gaaatttttc gccaagctga cctcagtgct
     4561 gaatacaatc ctgcagatgc ggaactcaaa gaccgggaca gaactggact atctgattag
     4621 ccccgtggct gatgtcaacg gaaacttctt cgacagcaga caggcaccca aaaatatgcc
     4681 tcaggatgca gacgccaacg gggcctacca catcgggctg aagggactga tgctgctggg
     4741 ccggatcaag aacaatcagg aggggaagaa gctgaacctg gtcattaaga acgaggaata
     4801 cttcgagttt gtccagaata gaaataacaa aaggccggcg gccacgaaaa aggccggcca
     4861 ggcaaaaaag aaaaagggat cctacccata cgatgttcca gattacgctt atccctacga
     4921 cgtgcctgat tatgcatacc catatgatgt ccccgactat gcctaagaat tctgcagata
     4981 tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     5041 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5101 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5161 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5221 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     5281 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     5341 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5401 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5461 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5521 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5581 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5641 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5701 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     5761 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5821 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5881 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5941 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6001 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     6061 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     6121 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     6181 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     6241 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     6301 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     6361 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6421 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6481 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6541 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6601 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6661 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6721 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6781 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6841 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6901 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6961 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     7021 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     7081 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     7141 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     7201 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     7261 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7321 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     7381 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7441 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7501 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7561 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7621 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7681 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7741 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7801 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7861 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7921 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7981 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8041 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8101 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8161 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8221 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8281 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8341 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8401 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8461 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8521 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8581 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8641 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8701 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8761 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8821 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8881 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8941 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9001 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9061 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9121 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9181 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9241 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9301 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9361 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9421 ttccccgaaa agtgccacct gacgtc
//
