LOCUS       sequence-454883-        6428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SYNZIP18-Cascade..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6428)
  AUTHORS   Villegas Kcam MC, Tsong AJ, Chappell J
  TITLE     Uncovering the Distinct Properties of a Bacterial Type I-E CRISPR 
            Activation System.
  JOURNAL   ACS Synth Biol. 2022 Jan 25. doi: 10.1021/acssynbio.1c00496.
  PUBMED    35077145
REFERENCE   2  (bases 1 to 6428)
  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: "ACS Synth 
            Biol. 2022 Jan 25. doi: 10.1021/acssynbio.1c00496."
COMMENT     Sequence Label: pJEC619
FEATURES             Location/Qualifiers
     source          1..6428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      23..94
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      110..137
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(165..182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(299..844)
                     /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(379..398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(958..1052)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1073..1732)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1647..1666
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1733..1835)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     promoter        1865..1899
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     promoter        1865..1899
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     promoter        1865..1899
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1915..1926
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 tcgagtaagg atctccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
       61 tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc
      121 gggtgggcct ttctgcgttt atacctaggg atatattccg cttcctcgct cactgactcg
      181 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
      241 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
      301 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
      361 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
      421 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
      481 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
      541 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      601 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
      661 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
      721 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
      781 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
      841 tcaagaagat catcttatta atcagataaa atatttctag atttcagtgc aatttatctc
      901 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgttactag tgcttggatt
      961 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     1021 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc
     1081 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
     1141 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     1201 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     1261 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     1321 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     1381 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     1441 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     1501 tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
     1561 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
     1621 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
     1681 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
     1741 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     1801 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc cagatatcga
     1861 cgtctttacg gctagctcag tcctaggtat tatgctagcg aattcaaaag atctaaagag
     1921 gagaaaggat ctatgtctat cgcggccacc ctggaaaacg acttagcacg ccttgagaat
     1981 gaaaatgctc gtctggagaa agatatcgcg aatctcgagc gtgacctggc aaaattagag
     2041 cgtgaggaag cttatttcgg tggcggtggc tctggtggaa atttgcttat tgataactgg
     2101 atccctgtac gcccgcgaaa cggggggaaa gtccaaatca taaatctgca atcgctatac
     2161 tgcagtagag atcagtggcg attaagtttg ccccgtgacg atatggaact ggccgcttta
     2221 gcactgctgg tttgcattgg gcaaattatc gccccggcaa aagatgacgt tgaatttcga
     2281 catcgcataa tgaatccgct cactgaagat gagtttcaac aactcatcgc gccgtggata
     2341 gatatgttct accttaatca cgcagaacat ccctttatgc agaccaaagg tgtcaaagca
     2401 aatgatgtga ctccaatgga aaaactgttg gctggggtaa gcggcgcgac gaattgtgca
     2461 tttgtcaatc aaccggggca gggtgaagca ttatgtggtg gatgcactgc gattgcgtta
     2521 ttcaaccagg cgaatcaggc accaggtttt ggtggtggtt ttaaaagcgg tttacgtgga
     2581 ggaacacctg taacaacgtt cgtacgtggg atcgatcttc gttcaacggt gttactcaat
     2641 gtcctcacat tacctcgtct tcaaaaacaa tttcctaatg aatcacatac ggaaaaccaa
     2701 cctacctgga ttaaacctat caagtccaat gagtctatac ctgcttcgtc aattgggttt
     2761 gtccgtggtc tattctggca accagcgcat attgaattat gcgatcccat tgggattggt
     2821 aaatgttctt gctgtggaca ggaaagcaat ttgcgttata ccggttttct taaggaaaaa
     2881 tttaccttta cagttaatgg gctatggccc catccgcatt ccccttgtct ggtaacagtc
     2941 aagaaagggg aggttgagga aaaatttctt gctttcacca cctccgcacc atcatggaca
     3001 caaatcagcc gagttgtggt agataagatt attcaaaatg aaaatggaaa tcgcgtggcg
     3061 gcggttgtga atcaattcag aaatattgcg ccgcaaagtc ctcttgaatt gattatgggg
     3121 ggatatcgta ataatcaagc atctattctt gaacggcgtc atgatgtgtt gatgtttaat
     3181 caggggtggc aacaatacgg caatgtgata aacgaaatag tgactgttgg tttgggatat
     3241 aaaacagcct tacgcaaggc gttatatacc tttgcagaag ggtttaaaaa taaagacttc
     3301 aaaggggccg gagtctctgt tcatgagact gcagaaaggc atttctatcg acagagtgaa
     3361 ttattaattc ccgatgtact ggcgaatgtt aatttttccc aggctgatga ggtaatagct
     3421 gatttacgag acaaacttca tcaattgtgt gaaatgctat ttaatcaatc tgtagctccc
     3481 tatgcacatc atcctaaatt aataagcaca ttagcgcttg cccgcgccac gctatacaaa
     3541 catttacggg agttaaaacc gcaaggaggg ccatcaaatg gctgatgaaa ttgatgcaat
     3601 ggctttatat cgagcctggc aacaactgga taatggatca tgtgcgcaaa ttagacgtgt
     3661 ttcagaacct gatgaattac gcgatatccc tgcgttttat aggctggtgc aaccttttgg
     3721 ttgggaaaac ccacgtcacc agcaggctct tttgcgcatg gtgttttgcc tgagcgcagg
     3781 aaagaatgtc atccgacatc aggacaaaaa atcggagcaa acaacaggta tctcgttggg
     3841 aagagcttta gccaatagtg gaagaattaa cgagcgccgt atctttcaat taattcgggc
     3901 tgacagaaca gccgatatgg tccagttacg tcgattactt actcacgccg aacccgtact
     3961 tgactggcca ttaatggcca ggatgttgac ctggtgggga aagcgcgaac gccagcaact
     4021 tctggaagat tttgtattga ccacaaacaa aaatgcgtaa ggaaaccttt ctatgtctaa
     4081 ctttatcaat attcatgttc tgatctctca cagcccttca tgtctgaacc gcgacgatat
     4141 gaacatgcag aaagacgcta ttttcggcgg caaaagacga gtaagaattt caagtcaaag
     4201 ccttaaacgt gcgatgcgta aaagtggtta ttacgcacaa aatattggtg aatccagtct
     4261 cagaaccatt catcttgcac aattacgtga tgttcttcgg caaaaacttg gtgaacgttt
     4321 tgaccaaaaa atcatcgata agacattagc gctgctctcc ggtaaatcag ttgatgaagc
     4381 cgaaaagatt tctgccgatg cggttactcc ctgggttgtg ggagaaatag cctggttctg
     4441 tgagcaggtt gcaaaagcag aggctgataa tctggatgat aaaaagctgc tcaaagttct
     4501 taaggaagat attgccgcca tacgtgtgaa tttacagcag ggtgttgata ttgcgcttag
     4561 tggaagaatg gcaaccagcg gcatgatgac tgagttggga aaagttgatg gtgcaatgtc
     4621 cattgcgcat gcgatcacta ctcatcaggt tgattctgat attgactggt tcaccgctgt
     4681 agatgattta caggaacaag gttctgcaca tctgggaact caggaatttt catcgggtgt
     4741 tttttatcgt tatgccaaca ttaacctcgc tcaacttcag gaaaatttag gtggtgcctc
     4801 cagggagcag gctctggaaa ttgcaaccca tgttgttcat atgctggcaa cagaggtccc
     4861 tggagcaaaa cagcgtactt atgccgcttt taaccctgcg gatatggtaa tggttaattt
     4921 ctccgatatg ccactttcta tggcaaatgc ttttgaaaaa gcggttaaag cgaaagatgg
     4981 ctttttgcaa ccgtctatac aggcgtttaa tcaatattgg gatcgcgttg ccaatggata
     5041 tggtctgaac ggagctgctg cgcaattcag cttatctgat gtagacccaa ttactgctca
     5101 agttaaacaa atgcctactt tagaacagtt aaaatcctgg gttcgtaata atggcgaggc
     5161 gtgaacatga gatcttattt gatcttgcgg cttgctgggc caatgcaagc ctgggggcag
     5221 ccgacctttg aaggaacgcg acctaccgga agatttccga cccgaagcgg gttattaggg
     5281 ctactcgggg cttgtcttgg gatccaacgt gatgatactt cttcattaca ggcgttatca
     5341 gagagtgtgc aatttgcagt gcgctgcgat gaactcattc ttgacgatcg tcgtgtgtct
     5401 gtaacggggt tgcgtgatta ccatacagtc cttggagcgc gagaagatta ccgtggtttg
     5461 aaaagtcatg aaacgattca aacatggcgc gaatatttat gtgatgcctc ctttaccgtc
     5521 gctctctggt taacacccca tgcaacgatg gttatctcag aacttgaaaa agcagtatta
     5581 aagcctcggt atacacctta cctggggcgg agaagttgcc cactaacaca cccgcttttt
     5641 ttggggacat gtcaggcatc ggatcctcag aaggcgctat taaattatga gcccgttggc
     5701 ggcgatatat atagtgagga atcagttaca gggcatcatt taaaatttac ggcgcgcgac
     5761 gaaccgatga tcaccttgcc tcgacaattt gcttcccgag aatggtatgt gattaaagga
     5821 ggtatggatg tatctcagta aagtcatcat tgccagggcc tggagcaggg atctttacca
     5881 acttcaccag ggattatggc atttatttcc aaacagaccg gatgctgctc gtgattttct
     5941 ttttcatgtt gagaagcgaa acacaccaga aggctgtcat gttttattgc agtcagcgca
     6001 aatgcctgtt tcaactgccg ttgcgacagt cattaaaact aaacaggttg aatttcaact
     6061 tcaggttggt gttccactct attttcggct tcgggcaaat ccgatcaaaa ctattctcga
     6121 caatcaaaag cgcctggaca gtaaagggaa tattaaacgc tgtcgggttc cgttaataaa
     6181 agaagcagaa caaatcgcgt ggttgcaacg taaattgggc aatgcggcgc gcgttgaaga
     6241 tgtgcatccc atatcggaac ggccacagta tttttctggt gatggtaaaa gtggaaagat
     6301 ccaaacggtt tgctttgaag gtgtgctcac catcaacgac gcgccagcgt taatagatct
     6361 tgtacagcaa ggtattgggc cagctaaatc gatgggatgt ggcttgctat ctttggctcc
     6421 actgtaac
//
