LOCUS       sequence-486105-        3440 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Containing RP4 oriT and inc sequence, for eliminating RP4 plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3440)
  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 3440)
  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: DoS1
FEATURES             Location/Qualifiers
     source          1..3440
                     /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(754..840)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(909..1619)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1187..1206)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1456..1474
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             1626..1637
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(join(1647..1672,1673..1678,1679..1695,1696..1701,
                     1702..1720))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1677..1695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1702..1720
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     oriT            2319..2428
                     /note="incP origin of transfer"
     promoter        2476..2578
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2579..3238
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2645..2664)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3259..3353
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
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 acgccctagg tctagggcgg cggatttgtc ctactcagga gagcgttcac
      781 cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg
      841 atgcctctag attaattaat taagcggccg catcgatcgg gccctgaggc ctgcagggat
      901 ccaagctttt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt
      961 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg
     1021 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
     1081 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
     1141 ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt
     1201 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
     1261 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     1321 cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     1381 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
     1441 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     1501 ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     1561 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatg
     1621 gtacctttct cctctttaat gaattcggtc agtgcgtcct gctgatgtgc tcagtatctc
     1681 tatcactgat agggatgtca atctctatca ctgataggga ctcgaggtga agacgaaagg
     1741 gcctcgtgat acgcctagac agcccctcaa atgtcaatag gtgcgcccct catctgtcat
     1801 cactctgccc ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca
     1861 agtgtcaata ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg
     1921 cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa
     1981 atcgagcctg cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa
     2041 cgtccgcccc tcatctgtca gacgtcgata tctggcgaaa atgagacgtt gatcgtctct
     2101 cgcctgtccc ctcagttcag taatttcctg catttgcctg tttccagtcg gtagatattc
     2161 cacaaaacag cagggaagca gcgcttttcc gctgcataac cctgcttcgg ggtcattata
     2221 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     2281 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     2341 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     2401 gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg
     2461 atggctgtga gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag
     2521 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
     2581 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     2641 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     2701 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     2761 cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga
     2821 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     2881 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     2941 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     3001 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     3061 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     3121 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     3181 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
     3241 tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     3301 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca
     3361 ctagtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3421 gaaaagatca aaggatcttc
//
