LOCUS       sequence-379248-        8447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli - Eggerthella lenta shuttle plasmid (KanR), cumate-inducible
            promoter Pct5.1-crRNA(dadR-targeting spacer) dadR-deleting repair 
            template, used for dadR deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8447)
  AUTHORS   Dong X, Guthrie BGH, Alexander M, Noecker C, Ramirez L, Glasser NR, 
            Turnbaugh PJ, Balskus EP
  TITLE     Genetic manipulation of the human gut bacterium Eggerthella lenta 
            reveals a widespread family of transcriptional regulators.
  JOURNAL   Nat Commun. 2022 Dec 9;13(1):7624. doi: 10.1038/s41467-022-33576-3.
  PUBMED    36494336
REFERENCE   2  (bases 1 to 8447)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-12-9"; volume: "13(1):7624. doi"; pages: " 
            10.1038/s41467-022-33576-3"
COMMENT     Sequence Label: pXD71Cas10.1RT1-Pct5.1-crRNA(dadR)-RT(Delta-dadR)
FEATURES             Location/Qualifiers
     source          1..8447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      24..95
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(276..864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(356..375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3126..3144
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3301..3320
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             complement(3323..3934)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    4272..4299
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    4314..4341
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     terminator      4474..4501
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4624..5418)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 agcttgctgg ggcttttctt tttcaaataa aacgaaaggc tcagtcgaaa gactgggcct
       61 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gaggagctca tgccggcggc
      121 acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
      181 tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc
      241 gcacgcggcg catctcgggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
      721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1021 acttggtctg acagatggtt tccatccacg acatggcgaa ctgcgacacc atggtggtgg
     1081 cgaagtggcg cgggcccttg cagatctgcc aggcctcaac gttgttcggc gtttccagca
     1141 tgttcttcag gatggactcc gagtgcatgc accagatgcg cgacgtgccc acctggcagg
     1201 cgatggcctc gccgccgtgc ttcgccttga tgtccatgaa gttgtccacg atggactgca
     1261 tggcctcgtc ccacgagatg cgctgccagc cgggctcctc gcccttcggg ttcgtgcgct
     1321 tcatgggata gtgcagacgg tcggggtggt acgccgcctg gatggacgac tgcgacttcg
     1381 tgcagcagtt gcccatggac tggaacgctc cctcgtcgcc ctcgacgcgt atggcccgcc
     1441 cgttctggac gatgaccttg acgccgcact ccatcttgcc gcagccgcgg cagcacgttt
     1501 tcacggccac ggtgtcgttg ccggacacgc tgctgtacgt ctcttcggcc agcgcggtgt
     1561 gcgtcgaagc gccgaacgcc gcggctgccg caccggtgat ggcggcggtc ttgacgaacg
     1621 tgcgtcgtga catggtcagg ttacccatac tcctcctcct tctttctcct cgttctcgtc
     1681 ttgctcgtct tcgttcgggc gatcggggag gggggcttcc gggcgaagcc tcgggctccg
     1741 gaccggtgca aagtccccgt gtcgcttggg ttgaccatac ggatgcgagg ggatgcagag
     1801 catcgttcaa aggttggaaa gcgggttcga ccgcatcata aattcttatg atcttgtgat
     1861 catgaaaaaa tcgatgacga tttgaactgg aaaaacatca aacgaaggtc ataaactatt
     1921 ctgaaactag aataacttcc tcaaagaggg atgctcatct tttccgtata atcgaacccg
     1981 tcgacagagg ggtcggcgcg agcgacgttc cgtacgggct cacagggggt ggggcatgga
     2041 gacggcgtat ctgcacgagt tcgcgaaggt ggccgagtgc ggcagcttca cggcggctgc
     2101 gcgcgagctg catctgacgc agtcgacgct gagcaagcac gtggccttgt tggagcgcga
     2161 attcggggcc gacctgttcg tacgcgaccg cagcggcgtg agcctgaccg aggcgggcgg
     2221 cgtgctgtac gcgcaggcgc ggcagatgga gaagctgctg cgcgccaccg agctgctggt
     2281 gcgcgcggcg cgcgacgggc aggctgcggg ctcgactgcc ggtgtcgcgg cgggcgaggg
     2341 tgctgtcgac ggctcgcccg cggccgcggg ggacatggcg ctgcgctgca agtgccggct
     2401 cgcggccgag cgctgcgggc tcgaccggcg cgagctgggc gcgctcatcc tgtacgtgga
     2461 ggagcacggc ttcgagacga tccagggcga gctggcgctg acgcgcgacg aggtggccga
     2521 cgtgctgggc agcgtgtacc gcaagctggg cgtgggcgac aagcaggagg cgctcgattt
     2581 catccattcc gtttcggaat gatggtagac gcgcccgcgg cggttgggca tactgcgtag
     2641 aagtgaaagc gctatgaagc tggggggctt cgagcaaagg gcttcatccg aagaatggat
     2701 ggagaccgaa tatgagtgct tgtactgctg cgtccgtcga agacgcggcg aacaatcttc
     2761 ttgaggaact gaatagcggc gcgaagtgct tgtgcgtgga gcacgatgcg tccgacgacg
     2821 ttgcgccggc gttgctgcgc gcggctcgcg ctcgcgtcga ggcggggtcg aacgtgcgcg
     2881 tcgtgtgcgc cgacgccgtt gcagccgcgc gctttcgggc gcttgcggcc gacgatccgc
     2941 tgctgggcgc gctcgacgtg gtgtcggtgc gcgagctggc cctctcgatt ctgggcgatg
     3001 cgcgggtcgg cgacgccgtg ggacgcgacg cgcgcgtgct ggacgagaac gagctggcca
     3061 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3121 ataaaaatag gcgtatcacg aggccctttc gtcttcaagg cgaaaaaacc ccgccgaagc
     3181 ggggtttttt gcgaaatata atgaccctct tgataaccca agagggcatt ttttaggaac
     3241 ggttatttga ttattaatcc ggcggcaact gaaacgattc ggatcctgta ttactattct
     3301 gaattcgcgg ccgcttctag atctagcgct tgaatttcgc gtaccgctct cgcgcaattt
     3361 cgagtgtcga gttcctgaca cgctcaaagc gttctttgtc cttctgccat aggctacgaa
     3421 cagcaagccc acgcaccgaa ttgaatatca accaaaggat atcttctgca tcatcacgcg
     3481 aaagaccacg gctcaccaga acaccaagcc acatatcctc gacgacaaac cgattcctct
     3541 ctaccgtgcg ctgaataccc tcgcgtaacg ctgggtcccg gtcggcagcc acaatcaaat
     3601 caaggctgat agagaagtca tcgtcgagga aaaattcggc ggcgtcgtcc agcatttgct
     3661 ggatgacgtc atcctctggc ttcaatttcg ctaatcgagc ccgactgcgt tcggtgatct
     3721 gttcgtaaag ccattcaaaa gtggcaagca gaagctcaag ctttgtcggg aaatgatggc
     3781 tctgcgctcc tctggagaca ccagcagcac cgggcacatc tgcgatccgg aatcccgcgt
     3841 aacctttttc ccgtaaaacc cccagggccg ctgcaatcaa cttgccctgg gtctccattg
     3901 cgcgctctgc ctgtgttctt ctctttggac tcatgatcac catttttttt actaaagctt
     3961 gatctgcaat ttgaataata accactcctt tgtttatcca ccgaactaag ttggtgtttt
     4021 ttgaagcttg aattagatat ttaaaagtat catatctaat attataacta aattttctaa
     4081 aaaaaacatt gaaggaacgg ttatttgatt attaatccgg cttttttatt atttaggcaa
     4141 ctgaaacgat tcggatcctg tattactatt ctattttgtc aaaataattt tattgacaac
     4201 gtcttattaa cgttgatata atttaaattt tatttgacaa aaatgggctc gtgttgtaca
     4261 ataaatgtag taacaaacag acaatctggt ctgtttgtac gagtgttgcg gccaacaaac
     4321 agacaatctg gtctgtttgt attatgatta actaatgtca ctccccgcat ggggagtgcg
     4381 ggttgaaatg tctccgtcat cgtgcggcgc gagcgggtga gcggtcactc cccgcatggg
     4441 gagtgcgggt tgaaatacta gtcgactcac actggctcac cttcgggtgg gcctttctgc
     4501 gtttatatac tagagagaga atataaaaag ccagattatt aatccggctt ttttattatt
     4561 taattcctgt tataaaaaaa ggatcaattt tgaactctct cccaaagttg atcccttaac
     4621 gatctaaaac aattcatcca gtaaaatata atattttatt ttctcccaat caggcttgat
     4681 ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct ccctgatcga
     4741 ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc caagatcaat
     4801 aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt cgccgtggga
     4861 aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct cgcgcggatc
     4921 tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat cgttattcag
     4981 taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac aatccgatat
     5041 gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct tttcagggct
     5101 ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca tgagcagatt
     5161 gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct ttccttccag
     5221 ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat accggctgtc
     5281 cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct tagcaggaga
     5341 cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca attccggtga
     5401 tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt aaagataccc
     5461 caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt ctaaaacctt
     5521 aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa catagtatcg
     5581 acggagccga ttttgaaacc acaattatga tagaatttac aagctataag gttattgtcc
     5641 tgggtttcaa gcattagtcc atgcaagttt ttatgctttg cccattctat agatatattg
     5701 ataagaagac tggaatgccc gctccgccgg gattattcga aagcgcatcg tccaagcgag
     5761 accgtctgca ttcggaaatc tcggccggtt cgcgccgagg aagcccatct ggctgcatcc
     5821 caaatcgaca cacaacgcac ggaggccccg gcgatcgccg gggcctccga atcatgcgca
     5881 tcgtggaagc gaagctccgc aaaaaacccc gcttcagcgg ggttttttcg cgtcgcacct
     5941 ccttgtcgtg tattacaggt actactataa cacatcatct agcccgcccg cgatacgtcc
     6001 tctgcggtgc agcgtggtcg cggttccacg catccgccgc ccgctgcgca tcgcgcagca
     6061 cgtcattaag gctctgttgc cttggctgtt ccctgcgcat cagatattcg agcgggcgcg
     6121 atattcgact tctaagctct tccactttga ttcgcattcc cgcaactcct gctctgatgc
     6181 ggctcgcgag tccatcgcac ttaacaacga tgttatcgag gattgcgccg cgctcagctt
     6241 ttcccgcagc ttcccagccc cgaacctcgg tagcgatggg ttgaagctct ccaatgtctc
     6301 gcgcaactcc gtccgctcgt tgccgaagac actccaatcg gtcttgctca tcttcgatat
     6361 cgagcttcac gtcctcgagc tggctgtcaa gctcggcaag gtcaccctcc ttgtcggcaa
     6421 tctcggtaac aagctctgcc ttacgcgttt cggcatcatt cagtaaagcg gcggcttctt
     6481 ccctagccgc cgcgacgatt cgcgccgctt cgtccttcgc aggttcaacc acggccttat
     6541 cgaccgcaca gcgtaccttg tcgatatcca cggacttgtc cgtgtataca cgctgcgccc
     6601 gcgtctcctc gtcaagctcg attgcagggc ggtagcccat gcgcccctct aggtaatccc
     6661 caagctcctt gtgcatgctc cggtagaagc tgcgggggca catctgcttg tagttgaacc
     6721 gtccatctag gatgggcgtg aacggcacgt gcatgtggtc gcgggcgggc gtgccatcct
     6781 tcaacacctc gtccttgtgg acgtacccgc ccatcatgtt ctcgatgccg aacttcttcg
     6841 acaggtacca gtacgtgaga cggaagaacc tgtcctcatc gcccttgcgc acattgtcgg
     6901 gaagcgtcac aacaacgtca gccataacga ccgcatcctt gcgcgtggca cgcttccctg
     6961 cggccttctg ggcctcgttc acgcgctcga tgcggctctc caccgtgccc cagttcggct
     7021 cgccatcgcg cggctccacg cgccccacaa cgagcttgcc gtccttgttc gtgtagtggc
     7081 cgaccaccaa gtcgttcttg atgcgctcgg ggtcgatgtt gtcgcgctcc aggctgctgc
     7141 ggtcgcggcg gtagtgggca agcatgtgcc cgaccgatgg agccttgtat ttcgcgatgt
     7201 gagccatgga aacctccgaa ccgtacagac ttaatcgact aaagtctagc acgttagacc
     7261 ttagctacgc taaggacgtg gcagggccag ccctgcaccc gctcgggcgt tccccccgag
     7321 accccccagc ccttgcccgt tcgcggcttc gctcacggcc ctcgggccat cggcgcgagc
     7381 gccgaacgga gggacttcgc ccctccgaac cacccctcaa tctacctgcg gttcggcaca
     7441 cggcacacgg cactattagg ggggccgtcg tagtggcccc cctgtggtca tttgtcattt
     7501 gtcatttgtc acattatttt tcggacacat tatttttcgg acgtagttag cgaaaaataa
     7561 tgagccgccc gattctgcag gtctcatcca gtcgagacac gggcaaggaa agggtcgatg
     7621 gagcagagcg tccatcgacc cttaatcaag tctggtctat tcggttggcc gtcgcgtcaa
     7681 ttccgcaatc cgtgaacgag tcggcttaga tctgcgcgag cgcagaccac gccgccccgt
     7741 agtcacgcat tgctgcattg acccgactac tctttggcac ctgactgttc cacgcacttg
     7801 cgcttacgtt cggcgttcag atagttcccg cgacaatagc gctcgagaat cgagttgtac
     7861 cccacgatga tttcgcggta ctgctcggga aactccctca gcgccgcagc cgtgaaatcg
     7921 ttgatggtct cgaacctctc gcacatgatg aagccgacca actcgaagag catgtcctgc
     7981 ttgtccttag tcgagagctg ttcgagctga ccaatgtcga agttgtttcc ctcgatgcgt
     8041 gcttcgaccg gatacagctc cttgcccttc ttgcgacagg aagccgtgtc gtgaatcaga
     8101 tagtcatacg catgcctgat gttgttgatt ggctcggctg acgaacagca cttcttgccg
     8161 tccgcgctca atcggttcaa gacgttgatg attgccttgc gtgtcgtgtt gccgccccag
     8221 acaacaatga cgtgcgcatg cgtcttgcgc tgcttgctct tctggtcatg gtctatgtcg
     8281 tgaaccgcat aagcgaaagg cacttgaaga atgtcgtcta tctcgtcctg ccagtcatcg
     8341 cgcatgttct ctgggtagca gatgccctcc cagagcttca ccttcgtctc gccagcgttt
     8401 gccgccatgc tataatctcc ttgtctcgaa tgcccctgct gccgcca
//
