LOCUS       sequence-491752-        4141 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Containing RP4 oriT, RP4 inc and I-SceI enzyme sequence, for 
            eliminating RP4 plasmid and self-cutting..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4141)
  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 4141)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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
FEATURES             Location/Qualifiers
     source          1..4141
                     /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"
     promoter        1670..1725
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1671..1689
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1741..2367
                     /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     complement(1817..1836)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      complement(2389..2436)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     oriT            3020..3129
                     /note="incP origin of transfer"
     promoter        3177..3279
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3280..3939
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3346..3365)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3960..4054
                     /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 ccattaccct gttatcccta ttatttcagg aaagtttcgg aggagatagt gttcggcagt
      961 ttgtacatca tctgcgggat caggtacggt ttgatcaggt tgtagaagat caggtaagac
     1021 atagaatcga tgtagatgat cggtttgttt ttgttgattt ttacgtaaca gttcagttgg
     1081 aatttgttac gcagaccctt aaccaggtat tctacttctt cgaaagtgaa agactgggtg
     1141 ttcagtacga tcgatttgtt ggtagagttt ttgttgtaat cccatttacc accatcatcc
     1201 atgaaccagt atgccagaga catcggggtc aggtagtttt caaccaggtt gttcgggatg
     1261 gtttttttgt tgttaacgat gaacaggtta gccagtttgt tgaaagcttg gtgtttgaaa
     1321 gtctgggcgc cccaggtgat taccaggtta cccaggtggt taacacgttc ttttttgtgc
     1381 ggcggggaca gtacccactg atcgtacagc agacatacgt ggtccatgta tgctttgttt
     1441 ttccactcga actgcataca gtaggtttta ccttcatcac gagaacggat gtaagcatca
     1501 cccaggatca gaccgatacc tgcttcgaac tgttcgatgt tcagttcgat cagctgggat
     1561 ttgtattctt tcagcagttt agagttcgga cccaggttca ttacctggtt ttttttgatg
     1621 tttttcatat gcatataggt accctcctct ttagatcttt tgaattcttt tctctatcac
     1681 tgatagggag tggtaaaata actctatcaa cgatagagtg tcaacaaaaa ttaggaatta
     1741 atgatgtcta gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
     1801 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
     1861 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
     1921 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
     1981 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
     2041 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
     2101 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat
     2161 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
     2221 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
     2281 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
     2341 caacttaaat gtgaaagtgg gtcttaagca tgcctcgagg tgaagacgca aaaaacccct
     2401 caagacccgt ttagaggccc caaggggtta tgctagcctc gtgatacgcc taacgcgtga
     2461 cagcccctca aatgtcaata ggtgcgcccc tcatctgtca tcactctgcc cctcaagtgt
     2521 caaggatcgc gcccctcatc tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc
     2581 acttatcccc aggcttgtcc acatcatctg tgggaaactc gcgtaaaatc aggcgttttc
     2641 gccgatttgc gaggctggcc agctccacgt cgccggccga aatcgagcct gcccctcatc
     2701 tgtcaacgcc gcgccgggtg agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc
     2761 agacgtcgat atctggcgaa aatgagacgt tgatcgtctc tcgcctgtcc cctcagttca
     2821 gtaatttcct gcatttgcct gtttccagtc ggtagatatt ccacaaaaca gcagggaagc
     2881 agcgcttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
     2941 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
     3001 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
     3061 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
     3121 gctggccggc taccgccggc gtaacagatg agggcaagcg gatggctgtg agacgttgat
     3181 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     3241 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     3301 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     3361 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     3421 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     3481 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     3541 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     3601 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     3661 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     3721 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     3781 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     3841 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     3901 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg
     3961 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     4021 cggttgccgc cgggcgtttt ttattggtga gaatccaagc actagtcatg accaaaatcc
     4081 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4141 c
//
