LOCUS       sequence-124358-        8410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FnCpf1 and spacers 1-4 of CRISPR array under artificial 
            promoter. Cas genes are removed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8410)
  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 8410)
  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: pY002 (pFnCpf1_min)
FEATURES             Location/Qualifiers
     source          1..8410
                     /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"
     promoter        join(1426..1431,1432..1449,1450..1455)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        5373..5407
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             5746..6936
                     /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(6098..6117)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(7970..8410,1..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 tattttttac actttatgct
     1441 tccggctcgt atgttaggag gtctttatca tgtcaattta tcaagaattt gttaataaat
     1501 atagtttaag taaaactcta agatttgagt taatcccaca gggtaaaaca cttgaaaaca
     1561 taaaagcaag aggtttgatt ttagatgatg agaaaagagc taaagactac aaaaaggcta
     1621 aacaaataat tgataaatat catcagtttt ttatagagga gatattaagt tcggtttgta
     1681 ttagcgaaga tttattacaa aactattctg atgtttattt taaacttaaa aagagtgatg
     1741 atgataatct acaaaaagat tttaaaagtg caaaagatac gataaagaaa caaatatctg
     1801 aatatataaa ggactcagag aaatttaaga atttgtttaa tcaaaacctt atcgatgcta
     1861 aaaaagggca agagtcagat ttaattctat ggctaaagca atctaaggat aatggtatag
     1921 aactatttaa agccaatagt gatatcacag atatagatga ggcgttagaa ataatcaaat
     1981 cttttaaagg ttggacaact tattttaagg gttttcatga aaatagaaaa aatgtttata
     2041 gtagcaatga tattcctaca tctattattt ataggatagt agatgataat ttgcctaaat
     2101 ttctagaaaa taaagctaag tatgagagtt taaaagacaa agctccagaa gctataaact
     2161 atgaacaaat taaaaaagat ttggcagaag agctaacctt tgatattgac tacaaaacat
     2221 ctgaagttaa tcaaagagtt ttttcacttg atgaagtttt tgagatagca aactttaata
     2281 attatctaaa tcaaagtggt attactaaat ttaatactat tattggtggt aaatttgtaa
     2341 atggtgaaaa tacaaagaga aaaggtataa atgaatatat aaatctatac tcacagcaaa
     2401 taaatgataa aacactcaaa aaatataaaa tgagtgtttt atttaagcaa attttaagtg
     2461 atacagaatc taaatctttt gtaattgata agttagaaga tgatagtgat gtagttacaa
     2521 cgatgcaaag tttttatgag caaatagcag cttttaaaac agtagaagaa aaatctatta
     2581 aagaaacact atctttatta tttgatgatt taaaagctca aaaacttgat ttgagtaaaa
     2641 tttattttaa aaatgataaa tctcttactg atctatcaca acaagttttt gatgattata
     2701 gtgttattgg tacagcggta ctagaatata taactcaaca aatagcacct aaaaatcttg
     2761 ataaccctag taagaaagag caagaattaa tagccaaaaa aactgaaaaa gcaaaatact
     2821 tatctctaga aactataaag cttgccttag aagaatttaa taagcataga gatatagata
     2881 aacagtgtag gtttgaagaa atacttgcaa actttgcggc tattccgatg atatttgatg
     2941 aaatagctca aaacaaagac aatttggcac agatatctat caaatatcaa aatcaaggta
     3001 aaaaagacct acttcaagct agtgcggaag atgatgttaa agctatcaag gatcttttag
     3061 atcaaactaa taatctctta cataaactaa aaatatttca tattagtcag tcagaagata
     3121 aggcaaatat tttagacaag gatgagcatt tttatctagt atttgaggag tgctactttg
     3181 agctagcgaa tatagtgcct ctttataaca aaattagaaa ctatataact caaaagccat
     3241 atagtgatga gaaatttaag ctcaattttg agaactcgac tttggctaat ggttgggata
     3301 aaaataaaga gcctgacaat acggcaattt tatttatcaa agatgataaa tattatctgg
     3361 gtgtgatgaa taagaaaaat aacaaaatat ttgatgataa agctatcaaa gaaaataaag
     3421 gcgagggtta taaaaaaatt gtttataaac ttttacctgg cgcaaataaa atgttaccta
     3481 aggttttctt ttctgctaaa tctataaaat tttataatcc tagtgaagat atacttagaa
     3541 taagaaatca ttccacacat acaaaaaatg gtagtcctca aaaaggatat gaaaaatttg
     3601 agtttaatat tgaagattgc cgaaaattta tagattttta taaacagtct ataagtaagc
     3661 atccggagtg gaaagatttt ggatttagat tttctgatac tcaaagatat aattctatag
     3721 atgaatttta tagagaagtt gaaaatcaag gctacaaact aacttttgaa aatatatcag
     3781 agagctatat tgatagcgta gttaatcagg gtaaattgta cctattccaa atctataata
     3841 aagatttttc agcttatagc aaagggcgac caaatctaca tactttatat tggaaagcgc
     3901 tgtttgatga gagaaatctt caagatgtgg tttataagct aaatggtgag gcagagcttt
     3961 tttatcgtaa acaatcaata cctaaaaaaa tcactcaccc agctaaagag gcaatagcta
     4021 ataaaaacaa agataatcct aaaaaagaga gtgtttttga atatgattta atcaaagata
     4081 aacgctttac tgaagataag tttttctttc actgtcctat tacaatcaat tttaaatcta
     4141 gtggagctaa taagtttaat gatgaaatca atttattgct aaaagaaaaa gcaaatgatg
     4201 ttcatatatt aagtatagat agaggtgaaa gacatttagc ttactatact ttggtagatg
     4261 gtaaaggcaa tatcatcaaa caagatactt tcaacatcat tggtaatgat agaatgaaaa
     4321 caaactacca tgataagctt gctgcaatag agaaagatag ggattcagct aggaaagact
     4381 ggaaaaagat aaataacatc aaagagatga aagagggcta tctatctcag gtagttcatg
     4441 aaatagctaa gctagttata gagtataatg ctattgtggt ttttgaggat ttaaattttg
     4501 gatttaaaag agggcgtttc aaggtagaga agcaggtcta tcaaaagtta gaaaaaatgc
     4561 taattgagaa actaaactat ctagttttca aagataatga gtttgataaa actgggggag
     4621 tgcttagagc ttatcagcta acagcacctt ttgagacttt taaaaagatg ggtaaacaaa
     4681 caggtattat ctactatgta ccagctggtt ttacttcaaa aatttgtcct gtaactggtt
     4741 ttgtaaatca gttatatcct aagtatgaaa gtgtcagcaa atctcaagag ttctttagta
     4801 agtttgacaa gatttgttat aaccttgata agggctattt tgagtttagt tttgattata
     4861 aaaactttgg tgacaaggct gccaaaggca agtggactat agctagcttt gggagtagat
     4921 tgattaactt tagaaattca gataaaaatc ataattggga tactcgagaa gtttatccaa
     4981 ctaaagagtt ggagaaattg ctaaaagatt attctatcga atatgggcat ggcgaatgta
     5041 tcaaagcagc tatttgcggt gagagcgaca aaaagttttt tgctaagcta actagtgtcc
     5101 taaatactat cttacaaatg cgtaactcaa aaacaggtac tgagttagat tatctaattt
     5161 caccagtagc agatgtaaat ggcaatttct ttgattcgcg acaggcgcca aaaaatatgc
     5221 ctcaagatgc tgatgccaat ggtgcttatc atattgggct aaaaggtctg atgctactag
     5281 gtaggatcaa aaataatcaa gagggcaaaa aactcaattt ggttatcaaa aatgaagagt
     5341 attttgagtt cgtgcagaat aggaataact aattgacagc tagctcagtc ctaggtataa
     5401 tgctagcgct gatttaggca aaaacgggtc taagaacttt aaataatttc tactgttgta
     5461 gatgagaagt catttaataa ggccactgtt aaaagtctaa gaactttaaa taatttctac
     5521 tgttgtagat gctactattc ctgtgccttc agataattca gtctaagaac tttaaataat
     5581 ttctactgtt gtagatgtct agagcctttt gtattagtag ccggtctaag aactttaaat
     5641 aatttctact gttgtagatt agcgatttat gaaggtcatt tttttgtcta gctttaatgc
     5701 ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat
     5761 gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg
     5821 ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat
     5881 ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca
     5941 ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc
     6001 gactacgcga tcatggcgac cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg
     6061 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     6121 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     6181 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg
     6241 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     6301 aagggagagc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     6361 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     6421 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     6481 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     6541 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg
     6601 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc
     6661 attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc
     6721 aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc
     6781 ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca
     6841 tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg
     6901 cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta
     6961 acggattcac cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc
     7021 aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc
     7081 gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga
     7141 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
     7201 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
     7261 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcccctacg tgctgctgaa
     7321 gttgcccgca acagagagtg gaaccaaccg gtgataccac gatactatga ctgagagtca
     7381 acgccatgag cggcctcatt tcttattctg agttacaaca gtccgcaccg ctgtccggta
     7441 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     7501 tcatgcaact cgtaggacag gtgccggcag cgcccaacag tcccccggcc acggggcctg
     7561 ccaccatacc cacgccgaaa caagcgccct gcaccattat gttccggatc tgcatcgcag
     7621 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc taaccgtttt
     7681 tatcaggctc tgggaggcag aataaatgat catatcgtca attattacct ccacggggag
     7741 agcctgagca aactggcctc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     7801 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     7861 gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt
     7921 caggcgtagc accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc
     7981 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     8041 cacagacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     8101 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     8161 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     8221 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     8281 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     8341 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     8401 ttgccatacg
//
