LOCUS       sequence-378335-        2890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  short crRNA generator with spacer LEA2 with WT repeat region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2890)
  AUTHORS   Liu Y, Pinto F, Wan X, Yang Z, Peng S, Li M, Cooper JM, Xie Z, 
            French CE, Wang B
  TITLE     Reprogrammed tracrRNAs enable repurposing of RNAs as crRNAs and 
            sequence-specific RNA biosensors.
  JOURNAL   Nat Commun. 2022 Apr 11;13(1):1937. doi: 10.1038/s41467-022-29604-x.
  PUBMED    35410423
REFERENCE   2  (bases 1 to 2890)
  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: "Nat 
            Commun."; date: "2022-04-11"; volume: "13(1):1937. doi"; pages: " 
            10.1038/s41467-022-29604-x"
COMMENT     Sequence Label: pLY227
FEATURES             Location/Qualifiers
     source          1..2890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(31..42)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(159..186)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        194..228
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             240..251
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    1145..1166
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        1191..1293
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1294..1953
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1360..1379)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1974..2068
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2182..2727
                     /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     2628..2647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2844..2861
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggagaccaga aacaaaaaaa ggccgcgtta gcggccttca ataattgggc atagctctaa
       61 aacgaccgaa ctatataata acttctagtt ttattcgaaa gtaacaaacc attttcttgc
      121 gtaaacctgt acgatcctac aggtctctag tatataaacg cagaaaggcc cacccgaagg
      181 tgagccagtg tgattgacag ctagctcagt cctagggatt gtgctagcta ctagagatta
      241 aagaggagaa atactagatg aaaaacataa atgccgacga cacatacaga ataattaata
      301 aaattaaagc ttgtagaagc aataatgata ttaatcaatg cttatctgat atgactaaaa
      361 tggtacattg tgaatattat ttactcgcga tcatttatcc tcattctatg gttaaatctg
      421 atatttcaat cctagataat taccctaaaa aatggaggca atattatgat gacgctaatt
      481 taataaaata tgatcctata gtagattatt ctaactccaa tcattcacca attaattgga
      541 atatatttga aaacaatgct gtaaataaaa aatctccaaa tgtaattaaa gaagcgaaaa
      601 catcaggtct tatcactggg tttagtttcc ctattcatac ggctaacaat ggcttcggaa
      661 tgcttagttt tgcacattca gaaaaagaca actatataga tagtttattt ttacatgcgt
      721 gtatgaacat accattaatt gttccttctc tagttgataa ttatcgaaaa ataaatatag
      781 caaataataa atcaaacaac gatttaacca aaagagaaaa agaatgttta gcgtgggcat
      841 gcgaaggaaa aagctcttgg gatatttcaa aaatattagg ttgcagtgag cgtactgtca
      901 ctttccattt aaccaatgcg caaatgaaac tcaatacaac aaaccgctgc caaagtattt
      961 ctaaagcaat tttaacagga gcaattgatt gcccatactt taaaaattaa taacactgat
     1021 agtgctagtg tagatcacta ctagagctcg gtaccaaaga cgaacaataa gacgctgaaa
     1081 agcgtctttt ttcgttttgg tccctctagt attaattttg tcttcatgca tgaagacaac
     1141 taggctctag aagcggccgc gaattcagat atctggcgaa aatgagacgt tgatcggcac
     1201 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
     1261 ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     1321 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     1381 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     1441 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     1501 catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     1561 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     1621 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     1681 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
     1741 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     1801 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     1861 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     1921 cagtactgcg atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct
     1981 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     2041 ccgccgggcg ttttttattg gtgagaatcc aagcactagt aacaacttat atcgtatggg
     2101 gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat
     2161 ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     2221 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     2281 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     2341 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     2401 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     2461 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     2521 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     2581 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg
     2641 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     2701 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     2761 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     2821 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcccta
     2881 ggcctagtag
//
