LOCUS       Exported               10653 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses FnCpf1 locus including Cas genes and spacers 1-4 of CRISPR
            array..
ACCESSION   .
VERSION     .
KEYWORDS    pY001 (pFnCpf1_full)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10653)
  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 10653)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10653
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7989..9179
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(8341..8360)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(10213..219)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tattttgatg caatatttgt
     1441 cttaggtcat ccaagttatt atccaaaatt tggttttaaa ccagccacag aatatcagat
     1501 aaaatgtgaa tatgatgtcc cagcggatgt ttttatggta ctagatttgt cagctaaact
     1561 agctagttta aaaggacaaa ctgtctacta tgccgatgag tttggcaaaa ttttttagat
     1621 ctacaaaatt ataaactaaa taaagattct tataataact ttatatataa tcgaaatgta
     1681 gagaatttta taaggagtct ttatcatgtc aatttatcaa gaatttgtta ataaatatag
     1741 tttaagtaaa actctaagat ttgagttaat cccacagggt aaaacacttg aaaacataaa
     1801 agcaagaggt ttgattttag atgatgagaa aagagctaaa gactacaaaa aggctaaaca
     1861 aataattgat aaatatcatc agttttttat agaggagata ttaagttcgg tttgtattag
     1921 cgaagattta ttacaaaact attctgatgt ttattttaaa cttaaaaaga gtgatgatga
     1981 taatctacaa aaagatttta aaagtgcaaa agatacgata aagaaacaaa tatctgaata
     2041 tataaaggac tcagagaaat ttaagaattt gtttaatcaa aaccttatcg atgctaaaaa
     2101 agggcaagag tcagatttaa ttctatggct aaagcaatct aaggataatg gtatagaact
     2161 atttaaagcc aatagtgata tcacagatat agatgaggcg ttagaaataa tcaaatcttt
     2221 taaaggttgg acaacttatt ttaagggttt tcatgaaaat agaaaaaatg tttatagtag
     2281 caatgatatt cctacatcta ttatttatag gatagtagat gataatttgc ctaaatttct
     2341 agaaaataaa gctaagtatg agagtttaaa agacaaagct ccagaagcta taaactatga
     2401 acaaattaaa aaagatttgg cagaagagct aacctttgat attgactaca aaacatctga
     2461 agttaatcaa agagtttttt cacttgatga agtttttgag atagcaaact ttaataatta
     2521 tctaaatcaa agtggtatta ctaaatttaa tactattatt ggtggtaaat ttgtaaatgg
     2581 tgaaaataca aagagaaaag gtataaatga atatataaat ctatactcac agcaaataaa
     2641 tgataaaaca ctcaaaaaat ataaaatgag tgttttattt aagcaaattt taagtgatac
     2701 agaatctaaa tcttttgtaa ttgataagtt agaagatgat agtgatgtag ttacaacgat
     2761 gcaaagtttt tatgagcaaa tagcagcttt taaaacagta gaagaaaaat ctattaaaga
     2821 aacactatct ttattatttg atgatttaaa agctcaaaaa cttgatttga gtaaaattta
     2881 ttttaaaaat gataaatctc ttactgatct atcacaacaa gtttttgatg attatagtgt
     2941 tattggtaca gcggtactag aatatataac tcaacaaata gcacctaaaa atcttgataa
     3001 ccctagtaag aaagagcaag aattaatagc caaaaaaact gaaaaagcaa aatacttatc
     3061 tctagaaact ataaagcttg ccttagaaga atttaataag catagagata tagataaaca
     3121 gtgtaggttt gaagaaatac ttgcaaactt tgcggctatt ccgatgatat ttgatgaaat
     3181 agctcaaaac aaagacaatt tggcacagat atctatcaaa tatcaaaatc aaggtaaaaa
     3241 agacctactt caagctagtg cggaagatga tgttaaagct atcaaggatc ttttagatca
     3301 aactaataat ctcttacata aactaaaaat atttcatatt agtcagtcag aagataaggc
     3361 aaatatttta gacaaggatg agcattttta tctagtattt gaggagtgct actttgagct
     3421 agcgaatata gtgcctcttt ataacaaaat tagaaactat ataactcaaa agccatatag
     3481 tgatgagaaa tttaagctca attttgagaa ctcgactttg gctaatggtt gggataaaaa
     3541 taaagagcct gacaatacgg caattttatt tatcaaagat gataaatatt atctgggtgt
     3601 gatgaataag aaaaataaca aaatatttga tgataaagct atcaaagaaa ataaaggcga
     3661 gggttataaa aaaattgttt ataaactttt acctggcgca aataaaatgt tacctaaggt
     3721 tttcttttct gctaaatcta taaaatttta taatcctagt gaagatatac ttagaataag
     3781 aaatcattcc acacatacaa aaaatggtag tcctcaaaaa ggatatgaaa aatttgagtt
     3841 taatattgaa gattgccgaa aatttataga tttttataaa cagtctataa gtaagcatcc
     3901 ggagtggaaa gattttggat ttagattttc tgatactcaa agatataatt ctatagatga
     3961 attttataga gaagttgaaa atcaaggcta caaactaact tttgaaaata tatcagagag
     4021 ctatattgat agcgtagtta atcagggtaa attgtaccta ttccaaatct ataataaaga
     4081 tttttcagct tatagcaaag ggcgaccaaa tctacatact ttatattgga aagcgctgtt
     4141 tgatgagaga aatcttcaag atgtggttta taagctaaat ggtgaggcag agctttttta
     4201 tcgtaaacaa tcaataccta aaaaaatcac tcacccagct aaagaggcaa tagctaataa
     4261 aaacaaagat aatcctaaaa aagagagtgt ttttgaatat gatttaatca aagataaacg
     4321 ctttactgaa gataagtttt tctttcactg tcctattaca atcaatttta aatctagtgg
     4381 agctaataag tttaatgatg aaatcaattt attgctaaaa gaaaaagcaa atgatgttca
     4441 tatattaagt atagatagag gtgaaagaca tttagcttac tatactttgg tagatggtaa
     4501 aggcaatatc atcaaacaag atactttcaa catcattggt aatgatagaa tgaaaacaaa
     4561 ctaccatgat aagcttgctg caatagagaa agatagggat tcagctagga aagactggaa
     4621 aaagataaat aacatcaaag agatgaaaga gggctatcta tctcaggtag ttcatgaaat
     4681 agctaagcta gttatagagt ataatgctat tgtggttttt gaggatttaa attttggatt
     4741 taaaagaggg cgtttcaagg tagagaagca ggtctatcaa aagttagaaa aaatgctaat
     4801 tgagaaacta aactatctag ttttcaaaga taatgagttt gataaaactg ggggagtgct
     4861 tagagcttat cagctaacag caccttttga gacttttaaa aagatgggta aacaaacagg
     4921 tattatctac tatgtaccag ctggttttac ttcaaaaatt tgtcctgtaa ctggttttgt
     4981 aaatcagtta tatcctaagt atgaaagtgt cagcaaatct caagagttct ttagtaagtt
     5041 tgacaagatt tgttataacc ttgataaggg ctattttgag tttagttttg attataaaaa
     5101 ctttggtgac aaggctgcca aaggcaagtg gactatagct agctttggga gtagattgat
     5161 taactttaga aattcagata aaaatcataa ttgggatact cgagaagttt atccaactaa
     5221 agagttggag aaattgctaa aagattattc tatcgaatat gggcatggcg aatgtatcaa
     5281 agcagctatt tgcggtgaga gcgacaaaaa gttttttgct aagctaacta gtgtcctaaa
     5341 tactatctta caaatgcgta actcaaaaac aggtactgag ttagattatc taatttcacc
     5401 agtagcagat gtaaatggca atttctttga ttcgcgacag gcgccaaaaa atatgcctca
     5461 agatgctgat gccaatggtg cttatcatat tgggctaaaa ggtctgatgc tactaggtag
     5521 gatcaaaaat aatcaagagg gcaaaaaact caatttggtt atcaaaaatg aagagtattt
     5581 tgagttcgtg cagaatagga ataactaatt cattcaagaa tatattaccc tgtcagttta
     5641 gcgactatta cctctttaat aatttgcagg ggaattattt tagtaatagt aatatacaca
     5701 agagttattg attatatgga aaattatatt tagataacat ggttaaatga ttttatattc
     5761 tgtccttact cgatatattt gcataatatc tatagtaatg cctcagatac tacatactat
     5821 tcatctagcc aaacaaaagg gcgcgatgct cataaaagta tcgataaagg aatctatagt
     5881 accaaaaaag atgacctgat cggtatcgat gttattaacc ataaatatgg tttggttggt
     5941 aaaattgatg tttttcataa agataagggc ttacttgtgg agagaaaaag gcaaatcaag
     6001 actatctatg atggctataa atatcagctt tatgcgcaat atttttgtct ccaagagatg
     6061 ggctatgatg tcaaagccat taaattttat tcgatggttg ataataaatc atacccaata
     6121 gctataccaa cttcagctga gttagaaaag tttgaaaaac atattcaaac aatcaagcaa
     6181 tataatccaa tggataactc atttaggcaa aatattgaaa agtgtaaatt ttgtatatat
     6241 gcaaacttat gtgataaaac ggacttgtag attatgttta gtaaaaatga tattgaatca
     6301 aagaatatag tttttgttaa tatttttgat ggagtgaaac ttagtctatc attggggaat
     6361 atagttataa aagataaaga aactgatgag gtgaaaacta agctttctgt tcataaagtt
     6421 cttgcattgt ttatcgtagg taatatgacg atgacctcgc aacttttaga gacctgtaag
     6481 aaaaatgcta tacagctagt ttttatgaaa aatagcttta gaccatatct atgttttggt
     6541 gatattgctg aggctaattt tttagctaga tataagcaat atagtgtagt tgagcaagat
     6601 ataagtttag caaggatttt tataacatca aagatacgca atcaacataa cttagtcaaa
     6661 agcctaagag ataaaactcc agagcagcaa gagatagtca aaaagaataa acagctaata
     6721 gcagagttag aaaatacaac aagcctagcg gagctaatgg gtatagaggg caatgttgcc
     6781 aaaaatttct tcaaaggatt ctatggacat ttagatagtt ggcaagggcg caaacctaga
     6841 ataaaacagg atccatataa tgttgtttta gacttgggct atagtatgtt gtttaatttt
     6901 gtagagtgtt ttttgcgact ttttggcttt gatttataca agggcttttg tcatcagact
     6961 tggtataagc gtaaatccct agtttgtgac tttgttgagc catttagatg tatagtggat
     7021 aaccaagtta gaaaatcatg gaatctcggg caattttctg tagaggattt tggttgcaaa
     7081 aatgagcagt tttatataaa aaaagataaa acaaaagact actcaaaaat actttttgcc
     7141 gagattatca gctacaagct agagatattt gaatatgtaa gagaatttta tcgtgccttt
     7201 atgcgaggca aagaaattgc agagtatcca atattttgtt atgaaactag gagggtgtat
     7261 gttgatagtc agttatgatt ttagtaataa taaagtacgt gcaaagtttg ccaaatttct
     7321 agaaagttat ggtgtacgtt tacaatattc ggtatttgag ctcaaatata gcaagagaat
     7381 gttagacttg attttagctg agatagaaaa taactatgta ccactattta caaatgctga
     7441 tagtgtttta atctttaatg ctccagataa agatgtgata aaatatggtt atgcgattca
     7501 tagagaacaa gaggttgttt ttatagacta aaaattgcaa accttagtct ttatgttaaa
     7561 ataactacta agttcttaga gatatttaaa aatatgactg ttgttatata tcaaaatgct
     7621 aaaaaaatca tagattttag gtcttttttt gctgatttag gcaaaaacgg gtctaagaac
     7681 tttaaataat ttctactgtt gtagatgaga agtcatttaa taaggccact gttaaaagtc
     7741 taagaacttt aaataatttc tactgttgta gatgctacta ttcctgtgcc ttcagataat
     7801 tcagtctaag aactttaaat aatttctact gttgtagatg tctagagcct tttgtattag
     7861 tagccggtct aagaacttta aataatttct actgttgtag attagcgatt tatgaaggtc
     7921 atttttttgt ctagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg
     7981 caccgtgtat gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc accctggatg
     8041 ctgtaggcat aggcttggtt atgccggtac tgccgggcct cttgcgggat atcgtccatt
     8101 ccgacagcat cgccagtcac tatggcgtgc tgctagcgct atatgcgttg atgcaatttc
     8161 tatgcgcacc cgttctcgga gcactgtccg accgctttgg ccgccgccca gtcctgctcg
     8221 cttcgctact tggagccact atcgactacg cgatcatggc gaccacaccc gtcctgtgga
     8281 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     8341 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
     8401 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
     8461 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
     8521 gcttcctaat gcaggagtcg cataagggag agcgtcgacc gatgcccttg agagccttca
     8581 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     8641 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     8701 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     8761 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     8821 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga
     8881 cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc
     8941 ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag
     9001 gatcgctcgc ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga
     9061 tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat
     9121 accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa
     9181 tggaagccgg cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt
     9241 cttgcggaga actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc
     9301 catctccagc agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg
     9361 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     9421 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     9481 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     9541 gaagtcccct acgtgctgct gaagttgccc gcaacagaga gtggaaccaa ccggtgatac
     9601 cacgatacta tgactgagag tcaacgccat gagcggcctc atttcttatt ctgagttaca
     9661 acagtccgca ccgctgccgg tagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     9721 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgcccaac
     9781 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgcc ctgcaccatt
     9841 atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta
     9901 ttaacgaagc gctaaccgtt tttatcaggc tctgggaggc agaataaatg atcatatcgt
     9961 caattattac ctccacgggg agagcctgag caaactggcc tcaggcattt gagaagcaca
    10021 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
    10081 tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt
    10141 catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact
    10201 gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
    10261 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
    10321 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
    10381 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
    10441 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
    10501 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
    10561 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
    10621 tatcaccagc tcaccgtctt tcattgccat acg
//
