LOCUS       sequence-488910-        2969 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  Bacterial expression of mCpol under control of the pTet promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2969)
  AUTHORS   Doherty EE, Adler BA, Yoon PH, Hsieh K, Loi K, Armbuster EG, Lahiri 
            A, Bolling CS, Wilcox XE, Akkati A, Iavarone AT, Pogliano J, Doudna 
            JA
  TITLE     A miniature CRISPR-Cas10 enzyme confers immunity by an inverse 
            signaling pathway.
  JOURNAL   bioRxiv [Preprint]. 2025 Apr 1:2025.03.28.646030. doi: 
            10.1101/2025.03.28.646030.
  PUBMED    40196485
REFERENCE   2  (bases 1 to 2969)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Apr 1:2025.03.28.646030. doi: 
            10.1101/2025.03.28.646030."
COMMENT     Sequence Label: pBA1739
FEATURES             Location/Qualifiers
     source          1..2969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /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     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1134..1205
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1221..1248
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1276..1293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1410..1955)
                     /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(1490..1509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2069..2163)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2184..2843)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2758..2777
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2844..2946)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc
      721 ttttaagaag gagatataca tatgtacata gcgatagatg gcgatgatgt ggggagaaaa
      781 ataacctcat catacttatc caactcagaa gagaggttga catacataag caacaaatta
      841 aatgatacca caaaaaaaat atctaaaatg cttttatcca atggatttga gataattttc
      901 caagccgcag atggtgttac agcgaaaaca gacaacgagg tcaatcttaa ctttgttttt
      961 gataagataa aatcctattc ctttgacgaa attacatttt cagctggtgt gggagcgaat
     1021 ctaagagaag cctatgttgc actgttaaat tctaaaagca acggtaagaa tatgatttct
     1081 atttacaaag acatattata aggatccaaa ctcgagtaag gatctccagg catcaaataa
     1141 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1201 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatacctagg
     1261 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     1321 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     1381 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     1441 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     1501 gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
     1561 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
     1621 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc
     1681 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc
     1741 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
     1801 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
     1861 agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag
     1921 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa
     1981 aatatttcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
     2041 tacgatataa gttgttacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
     2101 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     2161 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     2221 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     2281 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     2341 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     2401 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     2461 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     2521 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     2581 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     2641 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     2701 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     2761 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     2821 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     2881 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     2941 cgatcaacgt ctcattttcg ccagatatc
//
