LOCUS       sequence-390593-        9538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses CRISPR Cascade-Cas3 Type IC proteins for in vivo assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9538)
  AUTHORS   O'Brien RE, Bravo JPK, Ramos D, Hibshman GN, Wright JT, Taylor DW
  TITLE     Structural snapshots of R-loop formation by a type I-C CRISPR 
            Cascade.
  JOURNAL   Mol Cell. 2023 Mar 2;83(5):746-758.e5. doi: 
            10.1016/j.molcel.2023.01.024. Epub 2023 Feb 16.
  PUBMED    36805026
REFERENCE   2  (bases 1 to 9538)
  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"; doi: 
            "10.1016/j.molcel.2023.01.024"; journalName: "Mol Cell"; date: 
            "2023-03-2- 2"; volume: "83"; issue: "5"; pages: "746-758.e5"
COMMENT     Sequence Label: pBAD_TypeIC_Cascade_cas3_invivo
FEATURES             Location/Qualifiers
     source          1..9538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(166..1044)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1071..1355
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1278..1297
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             6936..6953
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7009..7026)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      7179..7265
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7357..7384
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        7403..7494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7495..7563,7564..8355)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7713..7732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8526..9114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9015..9034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9268..9285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9300..9440
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9366..9385)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(9526..9538,1..10)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
       61 gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
      121 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
      181 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
      241 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
      301 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
      361 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
      421 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
      481 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
      541 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
      601 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
      661 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
      721 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
      781 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
      841 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
      901 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
      961 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     1021 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa
     1081 ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac
     1141 caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac
     1201 aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca
     1261 cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg
     1321 acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc taacaggagg
     1381 aattaaccat ggatccgagc tcgagatctg cagcatgaca catggggctg ttaagacata
     1441 cggcatcaga ctgcgggtat ggggcgacta cgcctgcttc acgcggccgg agatgaaggt
     1501 ggaacgggtg tcatacgatg tcatgccccc ttccgccgca cggggcatcc ttgaggccat
     1561 ccactggaag ccggccatca ggtggattgt ggacaggatt catgtgctgc ggcccatcgt
     1621 cttcgacaac gtgcggcgca acgaggtcag ttcgaagata cccaagccca atccggcaac
     1681 ggccatgcgc gacaggaagc cgctctactt tctggtcgat gacggcagca accgccagca
     1741 acgggcggcg accctgctgc gcaacgtcga ctatgtcatc gaagcccact tcgaactgac
     1801 agacaaggcc ggagcggaag acaacgcggg caagcatctg gacatcttca ggcgtcgtgc
     1861 gcgcgccggg cagtcgttcc agcagccatg cctcggctgc cgcgagtttc cggcgtcgtt
     1921 cgaactgctg gagggggatg tgccgctctc gtgctacgcg ggcgagaagc gcgaccttgg
     1981 ctacatgctg ctcgacatcg acttcgaacg cgacatgacg ccgctgttct tcaaggccgt
     2041 catggaagac ggagtgataa cgcccccgtc gcgcacgtca ccggaggtga gggcatgatc
     2101 ctgcaggcat tgcatggcta ttaccagcgc atgagcgcag accccgatgc ggggatgccg
     2161 ccctacggga ccagcatgga gaacatctcg ttcgcgctgg tgctggatgc gaaagggacg
     2221 ctacggggca tcgaggactt gcgggaacag gaaggcaaga agctgcggcc ccgcaagatg
     2281 ctggttccta tcgcggaaaa gaaggggaac ggcatcaaac cgaacttcct gtgggagaac
     2341 accagttata ttctgggggt ggacgccaag ggcaagcagg aacgtacaga caagtgccac
     2401 gccgcgttca ttgcccacat caaggcctac tgtgacactg cagaccaaga ccttgccgcc
     2461 gttctgcagt tccttgaaca cggcgaaaaa gacctctccg cttttccggt ttcggaagag
     2521 gtcatcggct ccaatatcgt ctttcgcatc gagggcgaac cgggcttcgt ccatgagcgt
     2581 ccggctgcgc ggcaggcttg ggccaactgc ctgaaccggc gggagcaagg gctatgtggt
     2641 caatgcctca tcaccgggga gcgccagaag ccgattgccc aactccatcc gtcgatcaaa
     2701 ggcgggcgag acggcgttcg cggtgcacag gctgttgcct cgatagtgtc ctttaacaac
     2761 accgcctttg aatcttacgg gaaagaacag tcgatcaatg cacccgtcag tcaagaagcg
     2821 gcattctctt atgtcacagc cctgaattac ctgctcaacc cgtcaaatcg acagaaggtc
     2881 accatcgctg acgccaccgt ggtcttctgg gccgaacgca gcagcccggc agaggacatc
     2941 tttgcaggca tgttcgaccc gccttccacc acggcgaaac cggagtcctc gaacgggacg
     3001 ccgccggaag acagtgaaga aggcagccag cctgacacag ccagagacga cccccatgcc
     3061 gctgccagaa tgcacgacct gcttgtggcc atccgttccg gcaagcgcgc cacggacatc
     3121 atgcccgaca tggacgagtc cgtgcgcttc catgtgcttg gcctgtcgcc caacgccgcc
     3181 cgcctgtccg tccgcttctg ggaggtcgat accgtcggtc acatgctgga caaggtgggg
     3241 cggcactacc gtgaactcga gatcattccg caattcaaca acgagcagga gttcccgtct
     3301 ctttcaaccc tgctgcggca gactgccgtc ttgaacaaga ccgagaacat ctcgcctgta
     3361 cttgcaggcg ggctattcag ggccatgctc accggcgggc cctatccgca aagcctgttg
     3421 cccgccgtgc ttggacgcat ccgcgccgag cacgccagac cagaagataa aagccgctat
     3481 aggcttgaag ttgtgacgta ttatcgagcc gccttgatca aggcgtatct cataagaaac
     3541 agaaagctgg aggtgcccgt gtctctcgac cccgcaagaa cagaccgccc gtacctgctc
     3601 ggacgactct tcgccgttct ggaaaaagcg caggaagatg ccgtgcccgg tgcgaacgcc
     3661 accatcaagg accgctatct ggcctccgca tcggccaatc cggggcaggt attccacatg
     3721 ctgctcaaga atgcgtccaa ccacacggcc aaactccgca aggaccctga aagaaagggg
     3781 tcggccatcc actacgaaat catgatgcaa gagattatcg acaacatcag cgattttccg
     3841 gtgaccatgt cgtccgacga acaaggcctg ttcatgatcg gctactacca ccagcgcaag
     3901 gcccttttca ccaagaagaa caaggagaac tagccatgac cgccattgcc aacagatacg
     3961 agttcgtgct gcttttcgat gtggagaacg gcaaccccaa cggcgacccc gacgcgggca
     4021 acatgccgcg catcgacccc gagaccgggc atggtctggt caccgacgtg tgcctgaagc
     4081 gcaagatacg caaccatgtg gccctgacaa aagaaggtgc cgagcggttc aatatctata
     4141 ttcaggaaaa ggccattctc aacgaaaccc acgaacgcgc ctacacggcc tgcgacctga
     4201 aacccgaacc caagaaactg cccaagaagg tcgaagacgc caagagggtc accgactgga
     4261 tgtgcacgaa cttctacgac atccgcacgt tcggcgccgt catgaccaca gaggtcaact
     4321 gcgggcaggt gcgcggcccc gtgcagatgg cctttgccag aagtgtggaa cccgtcgtgc
     4381 cgcaagaggt gagcatcacc cgcatggcag tcaccacgaa ggcagaggcc gaaaagcagc
     4441 agggagacaa ccgcaccatg ggtcgcaagc acatcgtgcc ctacggcctg tatgtggcgc
     4501 acggcttcat ctccgcaccg ctggcagaga agacaggctt ctccgatgaa gacctgacgc
     4561 tgttctggga cgccctcgtc aacatgttcg agcacgaccg ttcggcggca cgcggcctca
     4621 tgagcagccg caaactcatc gtcttcaagc accagaacag gctcggcaac gcccccgccc
     4681 acaagttgtt cgacctcgtg aaggtcagcc gtgcagaggg ttcgtccggc cccgcccggt
     4741 cgtttgccga ctatgcggtg acagtggggc aggcccccga aggcgtagag gtcaaagaga
     4801 tgctgtaggt tcgaatggca aacttggctg ctactttcgc tgaagcattt ggggcccgcg
     4861 agtggggaaa agcagccggc ttattgcatg atgctggaaa ggctacagct cagtttactc
     4921 aacgtctgga agggcgtcca gttcgcgtca accactcaat ttgcggggcg cgtctggccc
     4981 aggaacaggg ctctacctgt ggccttttac tttcctacgc aattgcggga caccacgggg
     5041 ggcttccgga tggtggatta caggacgggc aactgcatca ccgtttgaaa catgaacgtc
     5101 ttccagccga cgtctcgccc ccaagcgtag atatccgccc tgacgtgctg aaacccccgt
     5161 tcacatgccg tccagagcac cctggcttct ccttgagctt cttcacccgc atgttatttt
     5221 cttgtcttac tgacgcggat ttccttgaca cggaggcgtt ttgtacaccg gaaaaagcgt
     5281 cagcacgtaa cggccgttca gcgttaggat tggtcgctct tcgcgatgct cttaacactc
     5341 atcttgatac ggtggagcgc aaggctctgc cctcccgcgt taatgatatt cgtaagacag
     5401 tcctgcatga ttgtcgtgcg cgcgcttccg aaacgccggg attgttctcg ctgaccgtgc
     5461 ctactggcgg tggaaagacg ttgagcagca tggcctttgc ccttgatcat gctgtcacgc
     5521 acgggcttcg ccgtgttatc tacgcaattc ccttcacgtc aattatcgag cagaatgcga
     5581 aagtcttttc cgacgtattt gggcaagata acgttttgga gcatcattgt aactatcgca
     5641 gcaaggacga accggaagag caagggtacg acaaatggcg tggcctggca gctgaaaact
     5701 gggacgcgcc tgtagtcgtg actaccaacg tgcaattctt tgagagtttg ttttctaatc
     5761 gtccgtcccg ttgtcgtaaa ctgcacaaca tcgctcgttc ggtaattgtc ttagatgaag
     5821 cgcaggctat ccccaccgag tacttggagc catgcctgta cgccctgaag gaacttgtgg
     5881 gacagtatgg gtgcaccgtc gttttatgta cagcgaccca gccagcagtt gatgacgcat
     5941 ccctgccaga gcgtgttcgt ttacaccacg ttcgcgagat catcgcggat ccccaacgtt
     6001 tatatacgga ccttaaacgc acagaagtaa cattggcagg tcgcttaact gatgccgctt
     6061 tagccgcccg tctggacgga catggtcaag tcttatgcat tgtgggtaca aagccgcagg
     6121 ctcaagccgt gttctctctt ctgcaagaac gtgagggagc ttttcacctt agtacaaaca
     6181 tgtatccaga gcatcgtcgc cgtgtgcttg gaacaattcg ccaacgtctg gccgaccgct
     6241 tgccttgtcg cgtagtctct actagtttaa ttgaagcagg ggtcgatgtg gacttccctg
     6301 tcgtgtatcg tgctatggca ggacttgact ctatcgctca agccgcgggg cgttgtaatc
     6361 gtgaggggcg tttgcctgaa cctggtcagg tcgttgtcta tgagccagaa aaacctgccc
     6421 gcatgccctg gatgcaacgc tgcgccagtc gtgcacagga aacgctgcgc acccttcctg
     6481 aagccgatcc tctgggtctg gaagctatcc gtcgttattt tggtcttatc tacgatgtcc
     6541 aggaacttga ccgtaaagat attttcaagc gcctgcgcgg gcaggtagac cgcgatatgg
     6601 tattcaaatt tcgtgagatt gcgaatgact ttcgcttcat tgatgatgaa ggaactgctc
     6661 tggtaattcc gacaggacca gaagtagagg acttagtacg ccgtctgcgt ggttgtgagt
     6721 tcccgcgccc ggttttgcgc aagttgcagc agtattccgt gacagtgcgt catcgtgaac
     6781 tggagaaatt acgtagcgca ggagcagtgg aaatgatcgg agatgcttat ccggtcctgc
     6841 gcaatttggc agcgtactcg gaggatatgg gtttatgtgt cgactccgta gaagtttggc
     6901 agccagaagg tttggttagc aatagcgccg tcgaccatca tcatcatcat cattgagttt
     6961 aaacggtctc cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     7021 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     7081 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     7141 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     7201 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     7261 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     7321 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     7381 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     7441 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     7501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     7621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     7681 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     7741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     7801 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     8041 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     8101 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     8161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     8221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     8281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     8341 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     8401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     8461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     8521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     8581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8641 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     8701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     9001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     9061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     9121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     9181 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     9241 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     9301 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     9361 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     9421 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     9481 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcat
//
