LOCUS       sequence-355193-        8056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of X. albilineans type I-C Cascade genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8056)
  AUTHORS   Wimmer F, Mougiakos I, Englert F, Beisel CL
  TITLE     Rapid cell-free characterization of multi-subunit CRISPR effectors 
            and transposons.
  JOURNAL   Mol Cell. 2022 Feb 16. pii: S1097-2765(22)00102-2. doi: 
            10.1016/j.molcel.2022.01.026.
  PUBMED    35216669
REFERENCE   2  (bases 1 to 8056)
  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: "Mol Cell. 
            2022 Feb 16. pii: S1097-2765(22)00102-2. doi: 
            10.1016/j.molcel.2022.01.026."
COMMENT     Sequence Label: pXalb_IC_Cascade
FEATURES             Location/Qualifiers
     source          1..8056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..40
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(733..1392)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1307..1326
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1393..1495)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1887..1904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2021..2566)
                     /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(2101..2120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2642..2661
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2752..2786
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     promoter        2898..2932
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             2948..2959
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6374..6445
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6461..6488
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      7005..7091
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7183..7210
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        7229..7320
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(7778..8056,1..177)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7865..7884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
       61 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      121 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
      181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      421 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tcaggcattt gagaagcaca
      541 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
      601 tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt
      661 catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact
      721 gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
      781 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
      841 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
      901 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
      961 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     1021 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     1081 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     1141 tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg
     1201 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     1261 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     1321 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     1381 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     1441 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc
     1501 tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac caggatttat
     1561 ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga
     1621 tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg
     1681 tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca
     1741 ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg
     1801 tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat
     1861 atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     1921 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     1981 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     2041 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     2101 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
     2161 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
     2221 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
     2281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
     2341 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
     2401 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
     2461 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
     2521 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
     2581 taatcagata aaatatttct aggctcatga gcccgaagtg gcgagcccga tcttccccat
     2641 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     2701 cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttatcatcga tttgacagct
     2761 agctcagtcc taggtataat actagtatag gttaatgtca attcaagttt tagagctagg
     2821 tacctagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     2881 gtgctttttt tgaagctctg acagctagct cagtcctagg tataatgcta gccccgggaa
     2941 aagatctaaa gaggagaaag gatctatgag ctacggagtc aggctccata tctggggcga
     3001 acgggcgctg tttacgcgtc cggaaatgaa agtagaacgg atgtcgtatg acatcatcac
     3061 gccatcagcg gcacgcggca ttctggaagc gatccactgg aagccatcga tccgctgggt
     3121 ggtggacagc atccaggtgc tcaagcccat ccgcttcgag tcgatccgcc gcaacgaagt
     3181 tggtggcaag ctgtcggccg ccagcgtcag caaggcgatc aaagccgggc gcaccgatgg
     3241 cttggtcagc tacgtcgatg aggatcggca acagcgcgca gccaccctgc tacgcgatgt
     3301 cggctacgtt atcgccgcgc acttcgagtt gaccgataag gctggccccg acgacaacgt
     3361 cggcaagcac ttggacatct tcaaccgccg cgcgcggcgc ggacaatgct tccaggcgcc
     3421 ttgtctgggc acccgcgaat tcccggccag cttctccctg ctcgaagctg acgatgcacc
     3481 accgcccagc gatccgtcgc tgggcggcga acgcgacctg ggctggatgc tgcacgacat
     3541 cgactttgcc gatggcatga ccccacgctt ctttcgcgca cacatggtgg atggtcgcgt
     3601 cgtggtgcca ccctcgcacg acgacgaggt gcacacatga ttctgtctgc cctcgccgat
     3661 ttctaccagc gattactgga cgatcccgaa tctggtatcg ctgcgcccgg ctacagccag
     3721 cagaacatcg gctacaccgt cgtgctggcc gccgacggtc gcgtcgtcga tgtgaatgac
     3781 gaacacagca ccgagggcaa aaaacggctc gccatggcga tgagcgtgcc acagccggaa
     3841 aagcgcactg cgggaatcaa gtccaacttc ttgtgggaca aaaccagcta cacactcggc
     3901 atcagcgcca gcagcaagcg cagcgagcag gaacatgccg ccttcatcgc cttacatcag
     3961 caagcgctgg ccggcagcga cgatcctggc ctgcgggcgt tgctggcgtt tctgcaaacg
     4021 tggacgcctg cgcagtatac agaccacccg cagttcgtca gtcatggcga ggccttgctg
     4081 gatgcgaaca tcgtgttcaa gctgcaaggc gacagcggtt atttgcatca acgcacagcc
     4141 gcgcgtgcgg cgtgggaccg acaacaaggc catggcactg atagccccga aggcatctgc
     4201 ctggtcagcg gcaggcacgc gccgctagcg cggctgcatc ctgccatcaa gggcgtgaac
     4261 ggtgcacaga gttccggcgc ttccatcgtt tcgttcaatc tcgaggcctt cacctcctac
     4321 ggcaaaagcc agggcgaaaa cgccccagtg tcagagcagg ccgccttcgc ctacaccacc
     4381 gcactcaatc acctgctgcg gcgcgacccg cataaccgtc agcgcgtgca gatcggcgac
     4441 accaccgtgg tgttctgggc gcaagcgaaa acggcggaac acgcacaaaa tgccgaggac
     4501 gtcttcgccg acttcatgcg cgtcgacgaa gctgacgacc ccagcatagc cgacggccag
     4561 gttacccgac gcctggagtt ggccctcgaa caggtaagac aggcactgcc cctgcgcgag
     4621 ttggacgaca cgctggatga cgacgcccgc attttcgtgc tcgggctggc acccaatgcc
     4681 tcgcgcctgt ccatccgctt ctgggagaca cagacactcc gccagttcgt caaacgccag
     4741 gcagaccact acgacgatct gacgctcgtc ccagcggcat ggaaacgccc gccaacgccc
     4801 gccttccttg ccctgcaaac cgcgcccgtt tacggcgagc atggcaaacc caaggccgaa
     4861 gacgtctcgc ccttgctcgc aggcgaattg acccggtcga ttcttgccgg cacacgctat
     4921 ccgggcagct tgctcggcgc gatcgtcatg cgcttccgcg ccgacggaca actcactcca
     4981 ttacgcattg ccctgtgccg agccgttttg gcacgccagg cgcgtttgaa cacgcaacag
     5041 ggactgccca ccgaccagaa ggaaccaccc atgagtcttg acacaagcaa caccgaccca
     5101 ggctacctac tcggacggct ttttttccta ttttataacg ctcaagagaa agcattgggc
     5161 gctgtaagct caaatataaa ggacaagtac tatggatctg catcaaccac tccaaggcta
     5221 gtcttccctt ccctaataaa aaacttccag aatcatattt caaagttaag aaagatgggc
     5281 gaaggtgaaa agcgcattgc cggtgctatc gagcgagatg tcgaagaaat cattagtcat
     5341 ctgaaaggtc catatccgcg atccttaaaa attgagagcc aaggattttt ctcgattggc
     5401 tactaccatc agtccaaagc gcgcttcgtc cgcaaggacg gccaggacgc ccacgacacc
     5461 acttccgaag gagagaacgt atgagcgcca ttgcccaccg ctacgagttc gtttacctgt
     5521 tcgacgtcac caacggcaat cccaacggcg atcccgacgc tggcaacctg ccgcgactgg
     5581 acccggaaac caaccgtggg ctcgtcaccg atgtggcgct caagcgcaag atccgcaact
     5641 acgtcgcgct ggaaaaggaa ggccaatccg gctatgcgat ctacatgcag gaaaagtcgg
     5701 tactcaataa tcagcacaag caggcttatg ctgcactggg catcgaacca gagaccaaga
     5761 agctacccaa ggacgaggct aaagcgcgcg aactcaccgc gtggatgtgc gccaatttct
     5821 tcgatgtacg cagcttcggt gccgtgatga ccaccgaggt caactccgga caagtgcgtg
     5881 gccctgtcca acttgccttc gccagttcgg tggagccggt gctgccgctg gaagtatcga
     5941 tcacccgcgt ggcggtcacc cacgagaaag acctcgaaaa agagcgcacc atgggccgca
     6001 agcacatcct gccatacggc ctgtaccgcg ctcacggctt catttccgca aaactggccg
     6061 aacgcacggg tttctccgag gaagacctac agttgctgtg gcgtgcgctc accaacctgt
     6121 tcgagcacga ccgctccgct gcgcgcggcg agatggccgc acgcaagctg atcgtgttca
     6181 aacacgaaca tccaatgggt aacgcgccag cacacatgct attcgacaag gtcaaggttg
     6241 agcgagacga aggttccgac aacggaccgg cacgtagctt cgccgactac cgcgtggtca
     6301 tcgatcgcga tctgccggct ggcattacct tgaccgagct gttctgataa ctcgagtaag
     6361 gatctccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     6421 ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc
     6481 tttctgcgtt tatacctagg gagacgtcgg cgctgcaggc atgcaagctt ggctgttttg
     6541 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     6601 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     6661 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     6721 actgccaggc atcaaataaa acgaaaggct cagtcgaaag tttataccta gggagacgtc
     6781 ggcgctgcag gcatgcaagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     6841 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     6901 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     6961 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     7021 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     7081 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     7141 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     7201 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
     7261 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7321 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7381 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     7441 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7501 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7561 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7621 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7681 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7741 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
     7801 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     7861 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     7921 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     7981 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     8041 cgccctttga cgttgg
//
