LOCUS       sequence-191841-        8485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LshCas13a locus into pACYC184 with nontargeting 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: pC010 LshCas13a locus with nontargeting spacer
FEATURES             Location/Qualifiers
     source          1..8485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(636..641,642..659,660..665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      703..1248
                     /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     1149..1168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1365..1382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1774..1876
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1877..2536
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1943..1962)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             complement(3570..4760)
                     /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     4389..4408
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(4902..4936)
                     /label=BBa_J23111
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.58"
     promoter        complement(4902..4936)
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     promoter        complement(4902..4936)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 gatcttctcg atgatcttat acaggctcat attgatgttc ttgaagatct cgtagatgct
       61 cttctttgtc tccagttcat cgttttcaat gatgcgcaga atgatggagc agtcgttcag
      121 ggtcttatta gtgtactcat cccggatgtc aatttcgatc tcttcctcgt tctcctgtcg
      181 cttgatctcg atctttttat cgtccttagt aatgccctgc ctgatggcct cgtcaatgat
      241 ctttttcttt gtgatcccca gtgctttcag cttctcgctc ttgccgtagg cctcgatata
      301 cagcaccact tcctctgtct ccaggaagtc gtcgttattc tcaattctaa tgatcccttc
      361 tttgcccttc agtttgaaca ggatatttcc tgcatgaaat ttcctagtga actccttcag
      421 aatattatcg tttttcttgt agttgatgta ctttctgatg aacttgttgt tgtcgatctt
      481 ctctttgttg ttgttttcgt tgatgttcag aatgtacttg ttcccgtcat aattccgctt
      541 gactttcacc tttcgcttga tcttgaagtc tttcttatct cgcacctcgt accaccgctt
      601 gtgtccgaac aggttcccca tggataagac ctcctaacat acgagccgga agcataaagt
      661 gtaaatctag aaatatttta tctgattaat aagatgatct tcttgagatc gttttggtct
      721 gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt
      781 tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa
      841 acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc
      901 aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac
      961 gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca
     1021 gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc
     1081 cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag
     1141 ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt
     1201 gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc
     1261 gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc
     1321 gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg
     1381 aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca gccttttttc
     1441 tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt aagccagtat
     1501 acactccgct agcgctgatg tccggcggtg cttttgccgt tacgcaccac cccgtcagta
     1561 gctgaacagg agggacagct gatagaaaca gaagccactg gagcacctca aaaacaccat
     1621 catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg aataaatacc
     1681 tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg ataccgggaa
     1741 gccctgggcc aacttttggc gaaaatgaca cgttgatcgg cacgtaagag gttccaactt
     1801 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     1861 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     1921 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     1981 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     2041 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat
     2101 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     2161 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     2221 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     2281 taaagggttt attgagaata tgtttttcgt gtcagccaat ccctgggtga gtttcaccag
     2341 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     2401 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     2461 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     2521 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggttgcta
     2581 cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa attcgacccg
     2641 gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg tctattgctg gtttaccggt
     2701 ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg ccagtttgct
     2761 caggctctcc ccgtggaggt aataattgac gatatgatca tttattctgc ctcccagagc
     2821 ctgataaaaa cggttagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     2881 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgcttgtttc ggcgtgggta
     2941 tggtggcagg ccccgtggcc gggggactgt tgggcgctgc cggcacctgt cctacgagtt
     3001 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     3061 aggagctacc ggcagcggtg cggactgttg taactcagaa taagaaatga ggccgctcat
     3121 ggcgttgact ctcagtcata gtatcgtggt atcaccggtt ggttccactc tctgttgcgg
     3181 gcaacttcag cagcacgtag gggacttccg cgtttccaga ctttacgaaa cacggaaacc
     3241 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     3301 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     3361 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
     3421 gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt
     3481 ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa
     3541 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg
     3601 cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg
     3661 ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa
     3721 gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc
     3781 tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc
     3841 ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg
     3901 tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     3961 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     4021 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     4081 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     4141 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgacgctct
     4201 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     4261 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     4321 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     4381 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     4441 ccggccacga tgcgtccggc gtagaggatc cacaggacgg gtgtggtcgc catgatcgcg
     4501 tagtcgatag tggctccaag tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg
     4561 gacagtgctc cgagaacggg tgcgcataga aattgcatca acgcatatag cgctagcagc
     4621 acgccatagt gactggcgat gctgtcggaa tggacgatat cccgcaagag gcccggcagt
     4681 accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg
     4741 agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa
     4801 ctaccgcatt aaagcttgtt ttagtcccct tcgatattgg ggtggagatt gctgttctac
     4861 caagtaatcc atagttttag tccccttcga tattggggtg ggctagcact atacctagga
     4921 ctgagctagc tgtcaatcac agggtatcgt tagtattctc gattttggtc agcagttcaa
     4981 tgatcagatt cttaatgtag tcactgttgt aggactccag ttccaggacg ctcactttct
     5041 tgggcttcat caggcgctcc agaatatcat tgttgccgat cagtttgaat ttctttttca
     5101 gctcatcgta gtccagattc acgtcctttt taaagacttc gaacacgctg gcataagttg
     5161 aattgttgta gcgtgtgcta tatgacagca ggttgctcac ccgatcgatc tgctcagcaa
     5221 tactgtagtc ggcaaatgga tttctcacga tgtagaaatg ggagatatag ttccgaatgc
     5281 tctcattttc aggcttgttg atctcggaat tttcgctcag gtcgattcca aacccgtagc
     5341 aaatcttctc gaacttcttg taggactctt catcaaagaa cttgtagtag gctgtggtag
     5401 tgtaaaagcc gtcgcttcca ttgcgctttg ggtatgccct gctgatcccg gtgttatagc
     5461 ctgacagctt aatgatgccc agctccctca gtccattcac gatatagtgc atatcccttt
     5521 cgaatcttgc catctgaata gccagcttcc agttgatgtc aatcaggtag gactcgattt
     5581 tgttcagata gttgaactcg accagatctc gaatcttttt gtactcagac acgcggttat
     5641 agtccttttc gaaggatttg tagttcttat tctggatgtt ctttgcaaag aaatcgtcgt
     5701 tctccttcag ctttttgatg tacttctcct tgtattcctt tgagtagccg ttcagtttat
     5761 cgttcagatt cttcagaata gcgtcgatct cactgatttt gttctttctg atgttctttc
     5821 catcgatatt aaacaggaat ttagcatcgg ccatcttgat gtcgttgcta atcaggccgt
     5881 agatcttgtc aaagttagga tttccgatgt tcaggaacag attctttttg tagatataca
     5941 gctcgttctt ccgttccttg ggatagtaga tctcctggaa cttgttctca ttttcagact
     6001 ccatatcctc aatcagattg tcgatttcct tcttgtactt tttcagaaag tcactgttga
     6061 aaatgatcct gcacaggatt ttagacttga tctcctgatc tttgtccttg atatactgat
     6121 ccaccttttt cttcaggtcc ttcttgttga tgttagacag ctttctctgt tcatcctgca
     6181 ggatgttgga tttcttgtcg atctcgaact tggtttcatc gtcgaagatg acaatcttct
     6241 ccagtttctt ttccagcaca tcacagccgt tgatatcgtc tttaaactcg gtcagaatgt
     6301 tgttcttgat gtcctcgtag tagttgttga agatttcttt cttagtctgg atcttgaagt
     6361 cgtcgaaatc cttttcgatc tctttcatct tctgaatgaa ctcttccagg ttcagattcc
     6421 agttctctgt gatgcattcg ttccgcaggg tattcagctg cataatctca tccaggatgt
     6481 caatgatatt ctggtattct gaggtgttca gccacacgct ggtggcgaag aatctgtttc
     6541 ggatcttatt aatcacggca ttactgttca gcagtgcaaa gatagaaatg atgtactcga
     6601 aatcgtcgtt gatgacaatg gttttgtcgc tggttttcac ggtgatccac tcataagtct
     6661 tgttatcgtt gatgtcctta atctgtttct tgatttcctg gatgttcatc ttgaagtctg
     6721 aaaagtcgaa cagctcttcg tagtttttgc gcagatatcc aatgtagcac tcgatcactt
     6781 tcttctgata tttcttgatt gccttgttgt tccccttact ggcagaaatc tgtgcgttct
     6841 tgtagtagtt ctcgatgatg ttctcatcga tttcgtcaat gttgcccagg gttttcttca
     6901 gttcctgcag gaagatattc ttggactcgt tctcttccag atcgtcctcc aggatcagtt
     6961 tcttatacag ctccttattg acgtagatca gagcgttcag cacaattttt tcagtctcga
     7021 ttgtgtcaaa aggctcattc ttgggattgt tgcggtacag gttcaggatt tcgggcagca
     7081 ctttgctgaa gctgggcaga tacttgatat cgttgttgtt ctcttcagag atcttgatgt
     7141 cattgatctt tgtgatgatg tttttcttgt ccttaaagac cacgtccagg ttcagagctt
     7201 tgctcacctc ttcatcggag atcttcagat tctgaatgat gttgatcact ttgttgtaat
     7261 cgtcctgagt cccctgcagg tccctctctt tggaaatggc atgcaggatg cggttccttt
     7321 cattagtgcc gatctttgta aactttcgaa tgaagttgtt tgtgatgttg ttcttgttat
     7381 cgatgaaatc caggtcgcgg atgatcttga tcttactatt caggattttc ttatccagca
     7441 catagttctt ctcccggtcg cctccaaaga aatcgatgtt ctcgtcgttg ttgatgtttt
     7501 ctcgggaaaa gatcttgttc agctccatat ttgtgctggc aaagaaggtg atcagttcca
     7561 ggtccagctc ttccttggcg tgcaggcggc tgaaatcgtc agtattcact gtggtcatat
     7621 cgatgtcgtt atgcctcagc ttccccaggt acataatgtg ctccagtgta tactgtttca
     7681 ctctcttcag aattttctct gacaggatac tttcattcag aatcttttcg atctcgatct
     7741 tctccatttt cttcagtctg accttctgct cgttcaccag gattttttca attctgccct
     7801 tcaggtatct gtagatgatc ttatacagct ccttctcttc atcagatttc ttggaaaatt
     7861 tcttgctgtc gaagttcacc ttgtaatgct tcttaaagat tccgaagatc tcggtatcac
     7921 agttcccttt cttcagttcc ttctccagtt tcttaatcag ctcgtcgatc ttgaactcag
     7981 ccaggatctt ctcaatcttt tccttgatag agttgttctt aatgttctcg acaaagaact
     8041 tcacgatctt atccttcttg ttttctctct cgattttaaa cttttcgtgc ttgtccagca
     8101 gcacatagga cttgatgtat gtccgatttc tccgcttctc caggaactcg ttattgactt
     8161 tcttcacttt ctcaatgcgc tttgtgatgt tccaaaattc cagctccttg atcacgaaat
     8221 cggcaatgtc ctcgacggtc agatccacat tgatgttcag aatcttttcc accagcattt
     8281 tcttattttt acttttcttt ttgtccccgc cgacgttcag gtaaaacttc acgaagccca
     8341 ggatctccag attagacttg atcttctccc ggatttccat gaagttggtc aggatcacgt
     8401 caatcttatc gtctttcagc agcttctccc tcaggtgctc ttcgtagtat ctattttcaa
     8461 acaccttttc tgtctcgttt tcgat
//
