LOCUS       sequence-124356-        8759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FnCpf1 and spacers 1-4 of CRISPR array. Cas genes are 
            removed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8759)
  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 8759)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
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."
COMMENT     Sequence Label: pY003 (pFnCpf1_delta Cas)
FEATURES             Location/Qualifiers
     source          1..8759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             6095..7285
                     /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(6447..6466)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(8319..8759,1..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 tatttctgtc tactatgccg
     1441 atgagtttgg caaaattttt tagatctaca aaattataaa ctaaataaag attcttataa
     1501 taactttata tataatcgaa atgtagagaa ttttataagg agtctttatc atgtcaattt
     1561 atcaagaatt tgttaataaa tatagtttaa gtaaaactct aagatttgag ttaatcccac
     1621 agggtaaaac acttgaaaac ataaaagcaa gaggtttgat tttagatgat gagaaaagag
     1681 ctaaagacta caaaaaggct aaacaaataa ttgataaata tcatcagttt tttatagagg
     1741 agatattaag ttcggtttgt attagcgaag atttattaca aaactattct gatgtttatt
     1801 ttaaacttaa aaagagtgat gatgataatc tacaaaaaga ttttaaaagt gcaaaagata
     1861 cgataaagaa acaaatatct gaatatataa aggactcaga gaaatttaag aatttgttta
     1921 atcaaaacct tatcgatgct aaaaaagggc aagagtcaga tttaattcta tggctaaagc
     1981 aatctaagga taatggtata gaactattta aagccaatag tgatatcaca gatatagatg
     2041 aggcgttaga aataatcaaa tcttttaaag gttggacaac ttattttaag ggttttcatg
     2101 aaaatagaaa aaatgtttat agtagcaatg atattcctac atctattatt tataggatag
     2161 tagatgataa tttgcctaaa tttctagaaa ataaagctaa gtatgagagt ttaaaagaca
     2221 aagctccaga agctataaac tatgaacaaa ttaaaaaaga tttggcagaa gagctaacct
     2281 ttgatattga ctacaaaaca tctgaagtta atcaaagagt tttttcactt gatgaagttt
     2341 ttgagatagc aaactttaat aattatctaa atcaaagtgg tattactaaa tttaatacta
     2401 ttattggtgg taaatttgta aatggtgaaa atacaaagag aaaaggtata aatgaatata
     2461 taaatctata ctcacagcaa ataaatgata aaacactcaa aaaatataaa atgagtgttt
     2521 tatttaagca aattttaagt gatacagaat ctaaatcttt tgtaattgat aagttagaag
     2581 atgatagtga tgtagttaca acgatgcaaa gtttttatga gcaaatagca gcttttaaaa
     2641 cagtagaaga aaaatctatt aaagaaacac tatctttatt atttgatgat ttaaaagctc
     2701 aaaaacttga tttgagtaaa atttatttta aaaatgataa atctcttact gatctatcac
     2761 aacaagtttt tgatgattat agtgttattg gtacagcggt actagaatat ataactcaac
     2821 aaatagcacc taaaaatctt gataacccta gtaagaaaga gcaagaatta atagccaaaa
     2881 aaactgaaaa agcaaaatac ttatctctag aaactataaa gcttgcctta gaagaattta
     2941 ataagcatag agatatagat aaacagtgta ggtttgaaga aatacttgca aactttgcgg
     3001 ctattccgat gatatttgat gaaatagctc aaaacaaaga caatttggca cagatatcta
     3061 tcaaatatca aaatcaaggt aaaaaagacc tacttcaagc tagtgcggaa gatgatgtta
     3121 aagctatcaa ggatctttta gatcaaacta ataatctctt acataaacta aaaatatttc
     3181 atattagtca gtcagaagat aaggcaaata ttttagacaa ggatgagcat ttttatctag
     3241 tatttgagga gtgctacttt gagctagcga atatagtgcc tctttataac aaaattagaa
     3301 actatataac tcaaaagcca tatagtgatg agaaatttaa gctcaatttt gagaactcga
     3361 ctttggctaa tggttgggat aaaaataaag agcctgacaa tacggcaatt ttatttatca
     3421 aagatgataa atattatctg ggtgtgatga ataagaaaaa taacaaaata tttgatgata
     3481 aagctatcaa agaaaataaa ggcgagggtt ataaaaaaat tgtttataaa cttttacctg
     3541 gcgcaaataa aatgttacct aaggttttct tttctgctaa atctataaaa ttttataatc
     3601 ctagtgaaga tatacttaga ataagaaatc attccacaca tacaaaaaat ggtagtcctc
     3661 aaaaaggata tgaaaaattt gagtttaata ttgaagattg ccgaaaattt atagattttt
     3721 ataaacagtc tataagtaag catccggagt ggaaagattt tggatttaga ttttctgata
     3781 ctcaaagata taattctata gatgaatttt atagagaagt tgaaaatcaa ggctacaaac
     3841 taacttttga aaatatatca gagagctata ttgatagcgt agttaatcag ggtaaattgt
     3901 acctattcca aatctataat aaagattttt cagcttatag caaagggcga ccaaatctac
     3961 atactttata ttggaaagcg ctgtttgatg agagaaatct tcaagatgtg gtttataagc
     4021 taaatggtga ggcagagctt ttttatcgta aacaatcaat acctaaaaaa atcactcacc
     4081 cagctaaaga ggcaatagct aataaaaaca aagataatcc taaaaaagag agtgtttttg
     4141 aatatgattt aatcaaagat aaacgcttta ctgaagataa gtttttcttt cactgtccta
     4201 ttacaatcaa ttttaaatct agtggagcta ataagtttaa tgatgaaatc aatttattgc
     4261 taaaagaaaa agcaaatgat gttcatatat taagtataga tagaggtgaa agacatttag
     4321 cttactatac tttggtagat ggtaaaggca atatcatcaa acaagatact ttcaacatca
     4381 ttggtaatga tagaatgaaa acaaactacc atgataagct tgctgcaata gagaaagata
     4441 gggattcagc taggaaagac tggaaaaaga taaataacat caaagagatg aaagagggct
     4501 atctatctca ggtagttcat gaaatagcta agctagttat agagtataat gctattgtgg
     4561 tttttgagga tttaaatttt ggatttaaaa gagggcgttt caaggtagag aagcaggtct
     4621 atcaaaagtt agaaaaaatg ctaattgaga aactaaacta tctagttttc aaagataatg
     4681 agtttgataa aactggggga gtgcttagag cttatcagct aacagcacct tttgagactt
     4741 ttaaaaagat gggtaaacaa acaggtatta tctactatgt accagctggt tttacttcaa
     4801 aaatttgtcc tgtaactggt tttgtaaatc agttatatcc taagtatgaa agtgtcagca
     4861 aatctcaaga gttctttagt aagtttgaca agatttgtta taaccttgat aagggctatt
     4921 ttgagtttag ttttgattat aaaaactttg gtgacaaggc tgccaaaggc aagtggacta
     4981 tagctagctt tgggagtaga ttgattaact ttagaaattc agataaaaat cataattggg
     5041 atactcgaga agtttatcca actaaagagt tggagaaatt gctaaaagat tattctatcg
     5101 aatatgggca tggcgaatgt atcaaagcag ctatttgcgg tgagagcgac aaaaagtttt
     5161 ttgctaagct aactagtgtc ctaaatacta tcttacaaat gcgtaactca aaaacaggta
     5221 ctgagttaga ttatctaatt tcaccagtag cagatgtaaa tggcaatttc tttgattcgc
     5281 gacaggcgcc aaaaaatatg cctcaagatg ctgatgccaa tggtgcttat catattgggc
     5341 taaaaggtct gatgctacta ggtaggatca aaaataatca agagggcaaa aaactcaatt
     5401 tggttatcaa aaatgaagag tattttgagt tcgtgcagaa taggaataac taattcattc
     5461 aagaatatat taccctgtca gtttagcgac tattacctct ttaataattt gcaggggaat
     5521 tattttagta atagtaatat acacaagagt tattgattat atggaaaatt atatttagat
     5581 aacatggtta aatgatttta tattctgtcc ttactcgata tattttttat agactaaaaa
     5641 ttgcaaacct tagtctttat gttaaaataa ctactaagtt cttagagata tttaaaaata
     5701 tgactgttgt tatatatcaa aatgctaaaa aaatcataga ttttaggtct ttttttgctg
     5761 atttaggcaa aaacgggtct aagaacttta aataatttct actgttgtag atgagaagtc
     5821 atttaataag gccactgtta aaagtctaag aactttaaat aatttctact gttgtagatg
     5881 ctactattcc tgtgccttca gataattcag tctaagaact ttaaataatt tctactgttg
     5941 tagatgtcta gagccttttg tattagtagc cggtctaaga actttaaata atttctactg
     6001 ttgtagatta gcgatttatg aaggtcattt ttttgtctag ctttaatgcg gtagtttatc
     6061 acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt
     6121 catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc
     6181 gggcctcttg cgggatatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct
     6241 agcgctatat gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg
     6301 ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat
     6361 catggcgacc acacccgtcc tgtggatcct ctacgccgga cgcatcgtgg ccggcatcac
     6421 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     6481 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     6541 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     6601 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     6661 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     6721 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     6781 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
     6841 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
     6901 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
     6961 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
     7021 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
     7081 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat
     7141 cattggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
     7201 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
     7261 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
     7321 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
     7381 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc
     7441 agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta
     7501 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga
     7561 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc
     7621 cgtgtttcgt aaagtctgga aacgcggaag tcccctacgt gctgctgaag ttgcccgcaa
     7681 cagagagtgg aaccaaccgg tgataccacg atactatgac tgagagtcaa cgccatgagc
     7741 ggcctcattt cttattctga gttacaacag tccgcaccgc tgtccggtag ctccttccgg
     7801 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     7861 gtaggacagg tgccggcagc gcccaacagt cccccggcca cggggcctgc caccataccc
     7921 acgccgaaac aagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
     7981 ctaccctgtg gaacacctac atctgtatta acgaagcgct aaccgttttt atcaggctct
     8041 gggaggcaga ataaatgatc atatcgtcaa ttattacctc cacggggaga gcctgagcaa
     8101 actggcctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     8161 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt
     8221 cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca
     8281 ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca
     8341 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acagacggca
     8401 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     8461 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     8521 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     8581 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     8641 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     8701 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacg
//
