LOCUS       sequence-486104-        2789 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  DoS1 derived plasmid without RP4 oriT and inc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2789)
  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 2789)
  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-Delta-inc-Delta-oriT
FEATURES             Location/Qualifiers
     source          1..2789
                     /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"
     primer_bind     complement(1741..1759)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1825..1927
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1928..2587
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1994..2013)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2608..2702
                     /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 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     1801 cgatatctgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact ttcaccataa
     1861 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     1921 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     1981 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     2041 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     2101 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa
     2161 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     2221 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     2281 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     2341 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     2401 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     2461 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     2521 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     2581 ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta atgacctcag
     2641 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
     2701 atccaagcac tagtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2761 gaccccgtag aaaagatcaa aggatcttc
//
