LOCUS       sequence-191844-        8485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LshCas13a locus into pACYC184 with targeting spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8485)
  AUTHORS   Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung 
            J, Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, 
            Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung DT, Sabeti PC, 
            Collins JJ, Zhang F
  TITLE     Nucleic acid detection with CRISPR-Cas13a/C2c2.
  JOURNAL   Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
  PUBMED    28408723
REFERENCE   2  (bases 1 to 8485)
  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: "Science. 
            2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321."
COMMENT     Sequence Label: pC009 LshCas13a locus with targeting spacer
FEATURES             Location/Qualifiers
     source          1..8485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(390..424)
                     /label=BBa_J23111
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.58"
     promoter        complement(390..424)
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     promoter        complement(390..424)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        complement(join(4609..4614,4615..4632,4633..4638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      4676..5221
                     /direction=RIGHT
                     /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     5122..5141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5338..5355
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5747..5849
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5850..6509
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5916..5935)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(join(7543..8485,1..248))
                     /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     8362..8381
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg
       61 agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga
      121 ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc
      181 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
      241 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
      301 agcttgtttt agtccccttc gatattgggg tggtatggat tacttggtag aacagcaatc
      361 tgttttagtc cccttcgata ttggggtggg ctagcactat acctaggact gagctagctg
      421 tcaatcacag ggtatcgtta gtattctcga ttttggtcag cagttcaatg atcagattct
      481 taatgtagtc actgttgtag gactccagtt ccaggacgct cactttcttg ggcttcatca
      541 ggcgctccag aatatcattg ttgccgatca gtttgaattt ctttttcagc tcatcgtagt
      601 ccagattcac gtccttttta aagacttcga acacgctggc ataagttgaa ttgttgtagc
      661 gtgtgctata tgacagcagg ttgctcaccc gatcgatctg ctcagcaata ctgtagtcgg
      721 caaatggatt tctcacgatg tagaaatggg agatatagtt ccgaatgctc tcattttcag
      781 gcttgttgat ctcggaattt tcgctcaggt cgattccaaa cccgtagcaa atcttctcga
      841 acttcttgta ggactcttca tcaaagaact tgtagtaggc tgtggtagtg taaaagccgt
      901 cgcttccatt gcgctttggg tatgccctgc tgatcccggt gttatagcct gacagcttaa
      961 tgatgcccag ctccctcagt ccattcacga tatagtgcat atccctttcg aatcttgcca
     1021 tctgaatagc cagcttccag ttgatgtcaa tcaggtagga ctcgattttg ttcagatagt
     1081 tgaactcgac cagatctcga atctttttgt actcagacac gcggttatag tccttttcga
     1141 aggatttgta gttcttattc tggatgttct ttgcaaagaa atcgtcgttc tccttcagct
     1201 ttttgatgta cttctccttg tattcctttg agtagccgtt cagtttatcg ttcagattct
     1261 tcagaatagc gtcgatctca ctgattttgt tctttctgat gttctttcca tcgatattaa
     1321 acaggaattt agcatcggcc atcttgatgt cgttgctaat caggccgtag atcttgtcaa
     1381 agttaggatt tccgatgttc aggaacagat tctttttgta gatatacagc tcgttcttcc
     1441 gttccttggg atagtagatc tcctggaact tgttctcatt ttcagactcc atatcctcaa
     1501 tcagattgtc gatttccttc ttgtactttt tcagaaagtc actgttgaaa atgatcctgc
     1561 acaggatttt agacttgatc tcctgatctt tgtccttgat atactgatcc acctttttct
     1621 tcaggtcctt cttgttgatg ttagacagct ttctctgttc atcctgcagg atgttggatt
     1681 tcttgtcgat ctcgaacttg gtttcatcgt cgaagatgac aatcttctcc agtttctttt
     1741 ccagcacatc acagccgttg atatcgtctt taaactcggt cagaatgttg ttcttgatgt
     1801 cctcgtagta gttgttgaag atttctttct tagtctggat cttgaagtcg tcgaaatcct
     1861 tttcgatctc tttcatcttc tgaatgaact cttccaggtt cagattccag ttctctgtga
     1921 tgcattcgtt ccgcagggta ttcagctgca taatctcatc caggatgtca atgatattct
     1981 ggtattctga ggtgttcagc cacacgctgg tggcgaagaa tctgtttcgg atcttattaa
     2041 tcacggcatt actgttcagc agtgcaaaga tagaaatgat gtactcgaaa tcgtcgttga
     2101 tgacaatggt tttgtcgctg gttttcacgg tgatccactc ataagtcttg ttatcgttga
     2161 tgtccttaat ctgtttcttg atttcctgga tgttcatctt gaagtctgaa aagtcgaaca
     2221 gctcttcgta gtttttgcgc agatatccaa tgtagcactc gatcactttc ttctgatatt
     2281 tcttgattgc cttgttgttc cccttactgg cagaaatctg tgcgttcttg tagtagttct
     2341 cgatgatgtt ctcatcgatt tcgtcaatgt tgcccagggt tttcttcagt tcctgcagga
     2401 agatattctt ggactcgttc tcttccagat cgtcctccag gatcagtttc ttatacagct
     2461 ccttattgac gtagatcaga gcgttcagca caattttttc agtctcgatt gtgtcaaaag
     2521 gctcattctt gggattgttg cggtacaggt tcaggatttc gggcagcact ttgctgaagc
     2581 tgggcagata cttgatatcg ttgttgttct cttcagagat cttgatgtca ttgatctttg
     2641 tgatgatgtt tttcttgtcc ttaaagacca cgtccaggtt cagagctttg ctcacctctt
     2701 catcggagat cttcagattc tgaatgatgt tgatcacttt gttgtaatcg tcctgagtcc
     2761 cctgcaggtc cctctctttg gaaatggcat gcaggatgcg gttcctttca ttagtgccga
     2821 tctttgtaaa ctttcgaatg aagttgtttg tgatgttgtt cttgttatcg atgaaatcca
     2881 ggtcgcggat gatcttgatc ttactattca ggattttctt atccagcaca tagttcttct
     2941 cccggtcgcc tccaaagaaa tcgatgttct cgtcgttgtt gatgttttct cgggaaaaga
     3001 tcttgttcag ctccatattt gtgctggcaa agaaggtgat cagttccagg tccagctctt
     3061 ccttggcgtg caggcggctg aaatcgtcag tattcactgt ggtcatatcg atgtcgttat
     3121 gcctcagctt ccccaggtac ataatgtgct ccagtgtata ctgtttcact ctcttcagaa
     3181 ttttctctga caggatactt tcattcagaa tcttttcgat ctcgatcttc tccattttct
     3241 tcagtctgac cttctgctcg ttcaccagga ttttttcaat tctgcccttc aggtatctgt
     3301 agatgatctt atacagctcc ttctcttcat cagatttctt ggaaaatttc ttgctgtcga
     3361 agttcacctt gtaatgcttc ttaaagattc cgaagatctc ggtatcacag ttccctttct
     3421 tcagttcctt ctccagtttc ttaatcagct cgtcgatctt gaactcagcc aggatcttct
     3481 caatcttttc cttgatagag ttgttcttaa tgttctcgac aaagaacttc acgatcttat
     3541 ccttcttgtt ttctctctcg attttaaact tttcgtgctt gtccagcagc acataggact
     3601 tgatgtatgt ccgatttctc cgcttctcca ggaactcgtt attgactttc ttcactttct
     3661 caatgcgctt tgtgatgttc caaaattcca gctccttgat cacgaaatcg gcaatgtcct
     3721 cgacggtcag atccacattg atgttcagaa tcttttccac cagcattttc ttatttttac
     3781 ttttcttttt gtccccgccg acgttcaggt aaaacttcac gaagcccagg atctccagat
     3841 tagacttgat cttctcccgg atttccatga agttggtcag gatcacgtca atcttatcgt
     3901 ctttcagcag cttctccctc aggtgctctt cgtagtatct attttcaaac accttttctg
     3961 tctcgttttc gatgatcttc tcgatgatct tatacaggct catattgatg ttcttgaaga
     4021 tctcgtagat gctcttcttt gtctccagtt catcgttttc aatgatgcgc agaatgatgg
     4081 agcagtcgtt cagggtctta ttagtgtact catcccggat gtcaatttcg atctcttcct
     4141 cgttctcctg tcgcttgatc tcgatctttt tatcgtcctt agtaatgccc tgcctgatgg
     4201 cctcgtcaat gatctttttc tttgtgatcc ccagtgcttt cagcttctcg ctcttgccgt
     4261 aggcctcgat atacagcacc acttcctctg tctccaggaa gtcgtcgtta ttctcaattc
     4321 taatgatccc ttctttgccc ttcagtttga acaggatatt tcctgcatga aatttcctag
     4381 tgaactcctt cagaatatta tcgtttttct tgtagttgat gtactttctg atgaacttgt
     4441 tgttgtcgat cttctctttg ttgttgtttt cgttgatgtt cagaatgtac ttgttcccgt
     4501 cataattccg cttgactttc acctttcgct tgatcttgaa gtctttctta tctcgcacct
     4561 cgtaccaccg cttgtgtccg aacaggttcc ccatggataa gacctcctaa catacgagcc
     4621 ggaagcataa agtgtaaatc tagaaatatt ttatctgatt aataagatga tcttcttgag
     4681 atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg
     4741 gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag
     4801 cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta
     4861 actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg
     4921 ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg
     4981 tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg
     5041 agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga
     5101 gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5161 ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa
     5221 aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg
     5281 aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc
     5341 gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt
     5401 gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat
     5461 agtaagccag tatacactcc gctagcgctg atgtccggcg gtgcttttgc cgttacgcac
     5521 caccccgtca gtagctgaac aggagggaca gctgatagaa acagaagcca ctggagcacc
     5581 tcaaaaacac catcatacac taaatcagta agttggcagc atcacccgac gcactttgcg
     5641 ccgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct ggtgtccctg
     5701 ttgataccgg gaagccctgg gccaactttt ggcgaaaatg acacgttgat cggcacgtaa
     5761 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
     5821 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     5881 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     5941 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     6001 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     6061 cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     6121 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     6181 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     6241 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtgtcagcc aatccctggg
     6301 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     6361 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     6421 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     6481 actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa
     6541 cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag
     6601 caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg
     6661 ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg
     6721 aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga tcatttattc
     6781 tgcctcccag agcctgataa aaacggttag cgcttcgtta atacagatgt aggtgttcca
     6841 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgcttgt
     6901 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc tgccggcacc
     6961 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     7021 cgcgcccacc ggaaggagct accggcagcg gtgcggactg ttgtaactca gaataagaaa
     7081 tgaggccgct catggcgttg actctcagtc atagtatcgt ggtatcaccg gttggttcca
     7141 ctctctgttg cgggcaactt cagcagcacg taggggactt ccgcgtttcc agactttacg
     7201 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     7261 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     7321 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc
     7381 caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt
     7441 tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc
     7501 ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg
     7561 gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat
     7621 ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg
     7681 tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg
     7741 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
     7801 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
     7861 gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     7921 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     7981 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     8041 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     8101 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     8161 cggtcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     8221 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     8281 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     8341 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     8401 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atccacagga cgggtgtggt
     8461 cgccatgatc gcgtagtcga tagtg
//
