LOCUS       sequence-375935-        7970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli - Eggerthella lenta shuttle plasmid (TetR), 
            N-3xFLAG-dadR(Delta-TM) complementation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7970)
  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 7970)
  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: pXD70Tet(DadRmt6)-N-3xFLAG-DadR(Delta-TM)
FEATURES             Location/Qualifiers
     source          1..7970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      366..393
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      5420..5491
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(5672..6260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5752..5771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6431..7222,7223..7291))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7054..7073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7292..7396)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7464..7482
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    7639..7658
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             7902..7967
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 attctacgta gcggccgcgg atccacggag accgacctca tcaaaatcgg catcggcgag
       61 gtcagcctga tgggcacggt gcccgaagag gccgccgacg acccggccga cccggtggat
      121 ccgtgcgcgc tgatcgcgga gcgcttcgcc ctgtcgccgc gcgagaccga cgtgcttccg
      181 ctgctgctgg agggccgcac catctcgcgc atccaggaga cgctgttcat ctcggcgggc
      241 acggtgagca cgcatatccg ccacatctac cagaagacgg gcgtggacaa tcgccaggag
      301 ctcatcgacc tggcccacga gacggccgcg caggacgagg cggtctgaac tagtcgactc
      361 acactggctc accttcgggt gggcctttct gcgtttatat actagagaga gaatataaaa
      421 agccagatta ttaatccggc ttttttatta tttaattcct gttataaaaa aaggatcaat
      481 tttgaactct ctcccaaagt tgatccctta acgatttaca ttaccttctg aaacatatgg
      541 cgcaccttgt ccaggcggct gtttggacgg cggggctgga tgaccggctg accgacagcg
      601 gcctgatatc ctttcagctc tgtaaggcat acgctccgcc cgttggtgta aaaggccaga
      661 tcagtacggt atgcctgtat acagcgggcg ggaatctcgc cagtaaagac aacttcatcc
      721 ttttttacct gggccgtttc gatggtggca cagtatttcg gtgcatcatg ataagccctg
      781 gaaaggtatt cctggggcgc atagaggatg aaggagagat aaggttccag cagctgcgtc
      841 cccgattcct tcaatgcctg ttccaataca atcggggcca atgagcggaa gtccgccggc
      901 gtgctgaccg gactgtaata aagcccgtat tcaaagcaaa tcttacagtc cgttacgttc
      961 cagccgaaca agccctgctc cagcccgtaa cggataccat ccctgacagc gttttgaaaa
     1021 ctctggttca agtatcccag cgaaacccgg ctctcgtatt gtacaccgga gccaagcgag
     1081 agtggtgtaa cagacagtcc tatggatgcc caaaacgggt tgggcggcac ctcgatatgg
     1141 atggtgtggc tggctgcttt gagcggccgc tccatataaa tgacggaggg ttcctttacc
     1201 actgtttcaa gcttgtattt ttccgacagc aaagcggaaa caacctccaa ctgcacccgg
     1261 cccaaaaaag aaagaatgat ctcatgggtg atggaatcca cttcgcaacg caaaagcggg
     1321 tcagtatccg caagttgcgt aagagcgtcc agcagccgtt ctctttgcgc tgccgttttc
     1381 ggcgcaatcg ccgtccgcag catggggagg gggtcctcgc gccacctttt acgagggagc
     1441 cgggtttggt cccctaatac atcgtttaac ctcacgctgt cgctgggaag gataacaatt
     1501 tcaccctgat aagcggtgtc tgtccgaaca atttcccctt tggatggaat acgcatctct
     1561 gtgattttca gcttttctct cccggccagg gccaccgtat cccgcaggcg cagcgttccg
     1621 ctgtataacc gtagatagac acgccgctgg ccgcaatcgg tgtactcaac cttgaaaacg
     1681 ctgccgcata gggcggcgcc cccctgttcc ccaatcggtt ggaacagccc tgtcaccgca
     1741 tccatcaacg gttgaatgcc aaggccattt ttggcgctgc catgatagac tgggaacagg
     1801 gaggcgtctt gaacccgctg ctgttcctcc cgcgcaagtt tttcccggct gattggttct
     1861 cctgcgatat acttttccaa taattcatcg ttattttcga tgaccgcatc ccatgcttct
     1921 atgtcggtat tttcctccag gactatttcc ggggacagcg acaccgtctg cttgatgata
     1981 atatcggcgg agagcttatc ccgaacagac tgaaccacgc tctgcaaatc aacgccagcc
     2041 tggtcgatct tgttgataaa gataacggtg ggaatgttca ttttccgcag ggcatggaac
     2101 agaatacggg tctgggcctg cacgccatct ttagcggaga tcaccaagat ggccccatct
     2161 aaaacagcca aagagcggta cacctccgcc aaaaaatcca tgtggccggg cgtatccaca
     2221 atgttaactt tacatctgtg ccactggaag gaagtgactg ccgcttgaat ggtaatccca
     2281 cgctgccgct ccaaaaacat ggtgtccgtc ctcgttgtcc ctttttcgac gctccccggt
     2341 tctgaaatgg ctccgctggc atatagcagg ctctccgtca aggtcgtctt tccagcgtct
     2401 acatgggcaa gaattccaat attgattatt ttcatgtgat tgtcctccct ttacagcccc
     2461 aaagggcata aaaatcccca gcagtaaaat acttttacca ctggggatta taagttgcgg
     2521 acatacacat atacagcata cacctgtttg tgattgctgt ttttggggat atgtcaaaat
     2581 tgataaggca aaagtattct taaattgggt acaaaaaact aagtccctac aaaaggagct
     2641 atcataatcc tttgttccca ctatttgatt atagttttat ttaagaatac cttgccgcat
     2701 attttttact ccttttctgg attaaatcat tgtatcacat cagttttagg aaagcaagta
     2761 cctaaaagaa atttttcttc cccttatatg taacaatcat accggcttcc tagcgttcag
     2821 aatgttttct gctgtctgct gtggtgcgca aaaaaccccg cttcagcggg gttttttcgc
     2881 gtcgcacctc cttgtcgtgt attacaggta ctactataac acatcatcta gcccgcccgc
     2941 gatacgtcct ctgcggtgca gcgtggtcgc ggttccacgc atccgccgcc cgctgcgcat
     3001 cgcgcagcac gtcattaagg ctctgttgcc ttggctgttc cctgcgcatc agatattcga
     3061 gcgggcgcga tattcgactt ctaagctctt ccactttgat tcgcattccc gcaactcctg
     3121 ctctgatgcg gctcgcgagt ccatcgcact taacaacgat gttatcgagg attgcgccgc
     3181 gctcagcttt tcccgcagct tcccagcccc gaacctcggt agcgatgggt tgaagctctc
     3241 caatgtctcg cgcaactccg tccactcgtt gccgaagaca ctccaatcgg tcttgctcat
     3301 cttcgatatc gagcttcacg tcctcgagct ggctgtcaag ctcggcaagg tcaccctcct
     3361 tgtcggcaat ctcggtaaca agctctgcct tacgcgtttc ggcatcattc agtaaagcgg
     3421 cggcttcttc cctagccgcc gcgacgattc gcgccgcttc gtccttcgca ggttcaacca
     3481 cggccttatc gaccgcacag cgtaccttgt cgatatccac ggacttgtcc gtgtatacac
     3541 gctgcgcccg cgtctcctcg tcaagctcga ttgcagggcg gtagcccatg cgcccctcta
     3601 ggtaatcccc aagctccttg tgcatgctcc ggtagaagct gcgggggcac atctgcttgt
     3661 agttgaaccg tccatctagg atgggcgtga acggcacgtg catgtggtcg cgggcgggcg
     3721 tgccatcctt caacacctcg tccttgtgga cgtacccgcc catcatgttc tcgatgccga
     3781 acttcttcga caggtaccag tacgtgagac ggaagaacct gtcctcatcg cccttgcgca
     3841 cattgtcggg aagcgtcaca acaacgtcag ccataacgac cgcatccttg cgcgtggcac
     3901 gcttccctgc ggccttctgg gcctcgttca cgcgctcgat gcggctctcc accgtgcccc
     3961 agttcggctc gccatcgcgc ggctccacgc gccccacaac gagcttgccg tccttgttcg
     4021 tgtagtggcc gaccaccaag tcgttcttga tgcgctcggg gtcgatgttg tcgcgctcca
     4081 ggctgctgcg gtcgcggcgg tagtgggcaa gcatgtgccc gaccgatgga gccttgtatt
     4141 tcgcgatgtg agccatggaa acctccgaac cgtacagact taatcgacta aagtctagca
     4201 cgttagacct tagctacgct aaggacgtgg cagggccagc cctgcacccg ctcgggcgtt
     4261 ccccccgaga ccccccagcc cttgcccgtt cgcggcttcg ctcacggccc tcgggccatc
     4321 ggcgcgagcg ccgaacggag ggacttcgcc cctccgaacc acccctcaat ctacctgcgg
     4381 ttcggcacac ggcacacggc actattaggg gggccgtcgt agtggccccc ctgtggtcat
     4441 ttgtcatttg tcatttgtca cattattttt cggacacatt atttttcgga cgtagttagc
     4501 gaaaaataat gagccgcccg attctgcagg tctcatccag tcgagacacg ggcaaggaaa
     4561 gggtcgatgg agcagagcgt ccatcgaccc ttaatcaagt ctggtctatt cggttggccg
     4621 tcgcgtcaat tccgcaatcc gtgaacgagt cggcttagat ctgcgcgagc gcagaccacg
     4681 ccgccccgta gtcacgcatt gctgcattga cccgactact ctttggcacc tgactgttcc
     4741 acgcacttgc gcttacgttc ggcgttcaga tagttcccgc gacaatagcg ctcgagaatc
     4801 gagttgtacc ccacgatgat ttcgcggtac tgctcgggaa actccctcag cgccgcagcc
     4861 gtgaaatcgt tgatggtctc gaacctctcg cacatgatga agccgaccaa ctcgaagagc
     4921 atgtcctgct tgtccttagt cgagagctgt tcgagctgac caatgtcgaa gttgtttccc
     4981 tcgatgcgtg cttcgaccgg atacagctcc ttgcccttct tgcgacagga agccgtgtcg
     5041 tgaatcagat agtcatacgc atgcctgatg ttgttgattg gctcggctga cgaacagcac
     5101 ttcttgccgt ccgcgctcaa tcggttcaag acgttgatga ttgccttgcg tgtcgtgttg
     5161 ccgccccaga caacaatgac gtgcgcatgc gtcttgcgct gcttgctctt ctggtcatgg
     5221 tctatgtcgt gaaccgcata agcgaaaggc acttgaagaa tgtcgtctat ctcgtcctgc
     5281 cagtcatcgc gcatgttctc tgggtagcag atgccctccc agagcttcac cttcgtctcg
     5341 ccagcgtttg ccgccatgct ataatctcct tgtctcgaat gcccctgctg ccgccaagct
     5401 tgctggggct tttctttttc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     5461 ttttatctgt tgtttgtcgg tgaacgctct ctactagagg agctcatgcc ggcggcacct
     5521 cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa
     5581 tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac
     5641 gcggcgcatc tcgggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5701 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5761 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5821 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5881 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5941 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6001 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6061 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6121 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6181 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6241 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6361 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6421 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6481 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6541 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6601 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6661 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6721 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
     6781 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6841 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6901 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6961 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7021 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     7081 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7141 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7201 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7261 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7321 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7381 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     7441 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaaggcg
     7501 aaaaaacccc gccgaagcgg ggttttttgc gaaatataat gaccctcttg ataacccaag
     7561 agggcatttt ttaggaacgg ttatttgatt attaatccgg cggcaactga aacgattcgg
     7621 atcctgtatt actattctga attcgcggcc gcttctagat gagcatcgtc caaaggttgg
     7681 aaagcaggtt cgaccgcatc ataaattctt atgatcttgt gatcatgaaa aattcgatag
     7741 tgctttgaac tggggaaact ttaaccgtag gtcataaatt attctgaaac tagaatagtt
     7801 tcctcaaaga gggccgctca tcttttccgt ataatcgaac ccgtcgacag aggggtcggc
     7861 gcgagcgacg ttatgtacgg gctcacaggg ggtggggcat ggactacaaa gaccatgacg
     7921 gtgattataa agatcatgac atcgattaca aggatgacga tgacaaggga
//
