LOCUS       sequence-486110-        4730 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Targeting pCU1 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4730)
  AUTHORS   Tsoi R, Son HI, Hamrick GS, Tang K, Bethke JH, Lu J, Maddamsetti R, 
            You L
  TITLE     A predatory gene drive for targeted control of self-transmissible 
            plasmids.
  JOURNAL   Sci Adv. 2025 Apr 4;11(14):eads4735. doi: 10.1126/sciadv.ads4735. 
            Epub 2025 Apr 2.
  PUBMED    40173243
REFERENCE   2  (bases 1 to 4730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2025-04-4"; volume: "11(14):eads4735. doi"; pages: " 
            10.1126/sciadv.ads4735. Epub 2025 Apr 2"
COMMENT     Sequence Label: DoS2-pCU1
FEATURES             Location/Qualifiers
     source          1..4730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(747..841)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(862..1521)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1436..1455
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1522..1624)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      2954..3001
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3023..3649)
                     /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     3554..3573
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        3665..3720
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    3701..3719
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      4550..4636
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgccctagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
      781 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
      841 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
      901 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
      961 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     1021 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     1081 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     1141 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     1201 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     1261 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     1321 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     1381 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     1441 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     1501 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     1561 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     1621 atcaacgtct caaccggtcg ctacttgtgt ataagagtca gctatatagg gacgtatcaa
     1681 agctacgaac gttcacacct tccaggccga agcgttcaga aactgcttta taaatttccg
     1741 ccctggaaac tcccagtgac atcatttctt ctatttccgg caacatcaac tgaagtgccg
     1801 ccctgatact tttgggcttc atagtaccct catttagaat gatgtaattt tgatgtattt
     1861 ctgatgtatt tatgatgtat ttttgatgta cttttgttgt actggctacc tcagtaccag
     1921 ataatgatgt aattctgttg tactgtcaac cttcaagtga tgtacttttg atgtaatttt
     1981 gatgtacttt tgttgtaata gtgttgtact ggtattcatg caagcatgaa tacggaatat
     2041 tgaacgcttt ttagtcagtt agtccttaac ctacgtagat cgctcctacc aaaaaagatg
     2101 attttttatc ccttaacctg ctatacacct aacgcagcgc ataaatcacg ctatcacact
     2161 gatttaaaaa taaaaaaata aatagcgtca atattcggta ataattaggt attaaaaaca
     2221 cctaataact cattgtttta taatgagttt gctttattac tttggtaata ctaagatttc
     2281 agcttgttag atcccaatct tttgtttaag aaatgcacaa tgtgaggact tgggaatccg
     2341 cccaccttgt tatgggaatt ggcccaccta tctatgggaa acaccccacc ttactatggg
     2401 aattagccca ccttgttatg ggaattggcc caccttgggc gaatttccac acagcaccgg
     2461 cgcccggcaa ggtgggcgga ttcccacacg gcaccgacgc ccggcaaggt gggcggattc
     2521 ccacacggca ccggcgcccg gcaacggtgg gcggatttcc acacagcacc ggcgcccggc
     2581 aaggtgggcg gattcccaca cggcaccggc gcccggcaag gtgggcggat tcccacacag
     2641 caccggcgcc cggcaaggtg ggcggatttc cacacagcac cggcgcccgg caaggtgggc
     2701 ggattcccac acggcaccgg cgcccggcaa ggtgggcgga ttcccacacg gcaccggcgc
     2761 ccggcaaggt gggcggattc ccacacggca ccggcgcccg gcaaggtggg cggatttcca
     2821 cacagcaccg gcgcccggca aggtgggcgg atttccacac agcaccggcg cccggcaagg
     2881 tgggcggatt tccacacggc accggcgccc ggcaaggtgg gcggatttcc aacgcgttag
     2941 gcgtatcacg aggctagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     3001 gcgtcttcac ctcgaggcat gcttaagacc cactttcaca tttaagttgt ttttctaatc
     3061 cgcatatgat caattcaagg ccgaataaga aggctggctc tgcaccttgg tgatcaaata
     3121 attcgatagc ttgtcgtaat aatggcggca tactatcagt agtaggtgtt tccctttctt
     3181 ctttagcgac ttgatgctct tgatcttcca atacgcaacc taaagtaaaa tgccccacag
     3241 cgctgagtgc atataatgca ttctctagtg aaaaaccttg ttggcataaa aaggctaatt
     3301 gattttcgag agtttcatac tgtttttctg taggccgtgt acctaaatgt acttttgctc
     3361 catcgcgatg acttagtaaa gcacatctaa aacttttagc gttattacgt aaaaaatctt
     3421 gccagctttc cccttctaaa gggcaaaagt gagtatggtg cctatctaac atctcaatgg
     3481 ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca atacaatgta ggctgctcta
     3541 cacctagctt ctgggcgagt ttacgggttg ttaaaccttc gattccgacc tcattaagca
     3601 gctctaatgc gctgttaatc actttacttt tatctaatct agacatcatt aattcctaat
     3661 ttttgttgac actctatcgt tgatagagtt attttaccac tccctatcag tgatagagaa
     3721 aagaattcaa aagatctaaa gaggagggta cctatatgca tatgaaaaac atcaaaaaaa
     3781 accaggtaat gaacctgggt ccgaactcta aactgctgaa agaatacaaa tcccagctga
     3841 tcgaactgaa catcgaacag ttcgaagcag gtatcggtct gatcctgggt gatgcttaca
     3901 tccgttctcg tgatgaaggt aaaacctact gtatgcagtt cgagtggaaa aacaaagcat
     3961 acatggacca cgtatgtctg ctgtacgatc agtgggtact gtccccgccg cacaaaaaag
     4021 aacgtgttaa ccacctgggt aacctggtaa tcacctgggg cgcccagact ttcaaacacc
     4081 aagctttcaa caaactggct aacctgttca tcgttaacaa caaaaaaacc atcccgaaca
     4141 acctggttga aaactacctg accccgatgt ctctggcata ctggttcatg gatgatggtg
     4201 gtaaatggga ttacaacaaa aactctacca acaaatcgat cgtactgaac acccagtctt
     4261 tcactttcga agaagtagaa tacctggtta agggtctgcg taacaaattc caactgaact
     4321 gttacgtaaa aatcaacaaa aacaaaccga tcatctacat cgattctatg tcttacctga
     4381 tcttctacaa cctgatcaaa ccgtacctga tcccgcagat gatgtacaaa ctgccgaaca
     4441 ctatctcctc cgaaactttc ctgaaataat agggataaca gggtaatgga tccctgcagg
     4501 cctcagggcc cgatcgatgc ggccgcttaa ttaattaatc tagaggcatc aaataaaacg
     4561 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     4621 cctgagtagg acaaatccgc cgccctagac ctagtcatga ccaaaatccc ttaacgtgag
     4681 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
