LOCUS       sequence-215715-        4543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas14b1 locus (including crRNA and tracrRNA) under control
            of a tetracycline inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4543)
  AUTHORS   Harrington LB, Burstein D, Chen JS, Paez-Espino D, Ma E, Witte IP, 
            Cofsky JC, Kyrpides NC, Banfield JF, Doudna JA
  TITLE     Programmed DNA destruction by miniature CRISPR-Cas14 enzymes.
  JOURNAL   Science. 2018 Oct 18. pii: science.aav4294. doi: 
            10.1126/science.aav4294.
  PUBMED    30337455
REFERENCE   2  (bases 1 to 4543)
  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. 
            2018 Oct 18. pii: science.aav4294. doi: 10.1126/science.aav4294."
COMMENT     Sequence Label: pLBH546_Tet-Cas14b1Locus
FEATURES             Location/Qualifiers
     source          1..4543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1616..1643
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1671..1688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1805..2350)
                     /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(1885..1904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2464..2558)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2579..3238)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3153..3172
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3239..3341)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(3371..3997)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     3902..3921
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        4013..4068
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    4049..4067
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             4086..4097
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 gcctgaatag cattcagaac cgtctgaact tttggaaaga actgctgtat aatccgagca
       61 accgcaacga aatcaaaatc aaagttgtga agtatgcccc taaaaccgat acacgtgaac
      121 atccgcacta ttatagcgaa gcagaaatta aaggtcgcat caaacgtctg gaaaagcagc
      181 tgaaaaagtt caaaatgccc aagtatccgg aatttaccag cgaaaccatt agcctgcagc
      241 gtgaactgta tagctggaaa aatccggatg aactgaaaat tagcagcatc accgataaaa
      301 acgagagcat gaactattac ggcaaagagt atctgaaacg ctatatcgat ctgatcaata
      361 gccagacacc gcagattctg ttagaaaaag aaaacaacag cttttatctg tgcttcccga
      421 tcaccaaaaa catcgaaatg ccgaaaatcg atgatacctt tgaaccggtt ggtattgatt
      481 ggggtattac ccgtaatatt gccgttgtta gcattctgga tagcaaaacc aaaaaaccga
      541 agttcgtgaa attctatagc gcaggttata ttctgggtaa acgcaaacac tataaaagcc
      601 tgcgtaaaca ctttggccag aaaaaacgtc aggacaaaat taacaaactg ggcaccaaag
      661 aagatcgctt cattgatagc aacattcaca aactggcctt tctgatcgtg aaagaaattc
      721 gcaatcatag caacaaaccg atcatcctga tggaaaacat taccgataat cgtgaagagg
      781 ccgaaaaaag catgcgtcag aacattctgc tgcatagcgt taaaagccgt ctgcagaact
      841 atattgcata taaagcactg tggaacaaca ttccgaccaa tctggttaaa ccggaacata
      901 ccagccagat ttgtaatcgt tgtggtcatc aggatcgcga aaaccgtccg aaaggtagca
      961 aactgtttaa atgcgtgaaa tgcaactata tgagcaacgc cgattttaac gccagcatta
     1021 atatcgcacg caaattttac atcggcgaat atgagccgtt ctacaaagat aacgagaaga
     1081 tgaaaagcgg tgtcaacagc attagcatgt aattttcatt cggcgaaggt ttgattatct
     1141 atatgatagt caaattgggc tgccgttgct acagccctta agataccttg ctcatcatag
     1201 ccgaaaaaga gcaactctcg aaagagagcg agtatcaatt tttaatttac ctcaaaaaaa
     1261 agaatgtctt tataaacagc catgcaaaaa catcagattc taattttgct ttcagaaact
     1321 ttgttttcca gccttaattc tttcgttcgc gaggcagttt ccgatattga gaaaatgaat
     1381 taaggaatgc aactacctta caccgcttgc gaaagttgtc agaagataat atttccgata
     1441 ttgagaaaat gaattaagga atgcaacaat tcagcttcca cagaatacaa cctatagctc
     1501 aacatcaaga actctgaact acacagcaac agatgcaaac ctcgataccg tctggtacca
     1561 gtacaatgga atcaacacaa cattatcagg caacatttcg ttctagagtc acactggctc
     1621 accttcgggt gggcctttct gcgtttatac ctagggatat attccgcttc ctcgctcact
     1681 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
     1741 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
     1801 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
     1861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg
     1921 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
     1981 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt
     2041 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2101 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
     2161 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga
     2221 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
     2281 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
     2341 acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt
     2401 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt tactagtgct
     2461 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     2521 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     2581 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     2641 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     2701 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     2761 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     2821 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     2881 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     2941 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     3001 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     3061 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     3121 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     3181 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     3241 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     3301 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaga
     3361 tatcgacgtc ttaagaccca ctttcacatt taagttgttt ttctaatccg catatgatca
     3421 attcaaggcc gaataagaag gctggctctg caccttggtg atcaaataat tcgatagctt
     3481 gtcgtaataa tggcggcata ctatcagtag taggtgtttc cctttcttct ttagcgactt
     3541 gatgctcttg atcttccaat acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat
     3601 ataatgcatt ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga ttttcgagag
     3661 tttcatactg tttttctgta ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac
     3721 ttagtaaagc acatctaaaa cttttagcgt tattacgtaa aaaatcttgc cagctttccc
     3781 cttctaaagg gcaaaagtga gtatggtgcc tatctaacat ctcaatggct aaggcgtcga
     3841 gcaaagcccg cttatttttt acatgccaat acaatgtagg ctgctctaca cctagcttct
     3901 gggcgagttt acgggttgtt aaaccttcga ttccgacctc attaagcagc tctaatgcgc
     3961 tgttaatcac tttactttta tctaatctag acatcattaa ttcctaattt ttgttgacac
     4021 tctatcgttg atagagttat tttaccactc cctatcagtg atagagaaaa gaattcaaaa
     4081 gatctaaaga ggagaaagga tctctttcgg cttgcgagcc gctgctggaa tatgattgcc
     4141 cagaaaacca tcaagattaa actgaatccg accaaagagc agatcatcaa actgaacagc
     4201 atcatcgaag agtacatcaa agtgagcaat ttcaccgcca aaaaaatcgc cgaaatccaa
     4261 gaaagcttta ccgatagcgg tctgacccag ggcacctgta gcgaatgtgg taaagaaaaa
     4321 acctaccgca aataccatct gctgaaaaaa gacaacaaac tgttctgcat cacgtgctac
     4381 aaacgtaaat atagccagtt taccctgcag aaggtggaat ttcagaataa aaccggtctg
     4441 cgtaatgttg caaaactgcc gaaaacctat tataccaatg caattcgttt tgccagcgat
     4501 acctttagcg gttttgatga aatcatcaag aaaaagcaga atc
//
