LOCUS       sequence-375918-        8894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli - Eggerthella lenta shuttle plasmid (TetR), 
            N-3xFLAG-dadR(Delta-DBD) complementation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8894)
  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 8894)
  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(DadRmt5)-N-3xFLAG-DadR(Delta-DBD)
FEATURES             Location/Qualifiers
     source          1..8894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(4987..5014)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6307..6372)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    6616..6635
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     primer_bind     complement(6792..6810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6878..6982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6983..7051,7052..7843)
                     /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     complement(7201..7220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8014..8602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8503..8522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(8783..8854)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttcgagacaa ggagattata gcatggcggc aaacgctggc gagacgaagg tgaagctctg
       61 ggagggcatc tgctacccag agaacatgcg cgatgactgg caggacgaga tagacgacat
      121 tcttcaagtg cctttcgctt atgcggttca cgacatagac catgaccaga agagcaagca
      181 gcgcaagacg catgcgcacg tcattgttgt ctggggcggc aacacgacac gcaaggcaat
      241 catcaacgtc ttgaaccgat tgagcgcgga cggcaagaag tgctgttcgt cagccgagcc
      301 aatcaacaac atcaggcatg cgtatgacta tctgattcac gacacggctt cctgtcgcaa
      361 gaagggcaag gagctgtatc cggtcgaagc acgcatcgag ggaaacaact tcgacattgg
      421 tcagctcgaa cagctctcga ctaaggacaa gcaggacatg ctcttcgagt tggtcggctt
      481 catcatgtgc gagaggttcg agaccatcaa cgatttcacg gctgcggcgc tgagggagtt
      541 tcccgagcag taccgcgaaa tcatcgtggg gtacaactcg attctcgagc gctattgtcg
      601 cgggaactat ctgaacgccg aacgtaagcg caagtgcgtg gaacagtcag gtgccaaaga
      661 gtagtcgggt caatgcagca atgcgtgact acggggcggc gtggtctgcg ctcgcgcaga
      721 tctaagccga ctcgttcacg gattgcggaa ttgacgcgac ggccaaccga atagaccaga
      781 cttgattaag ggtcgatgga cgctctgctc catcgaccct ttccttgccc gtgtctcgac
      841 tggatgagac ctgcagaatc gggcggctca ttatttttcg ctaactacgt ccgaaaaata
      901 atgtgtccga aaaataatgt gacaaatgac aaatgacaaa tgaccacagg ggggccacta
      961 cgacggcccc cctaatagtg ccgtgtgccg tgtgccgaac cgcaggtaga ttgaggggtg
     1021 gttcggaggg gcgaagtccc tccgttcggc gctcgcgccg atggcccgag ggccgtgagc
     1081 gaagccgcga acgggcaagg gctggggggt ctcgggggga acgcccgagc gggtgcaggg
     1141 ctggccctgc cacgtcctta gcgtagctaa ggtctaacgt gctagacttt agtcgattaa
     1201 gtctgtacgg ttcggaggtt tccatggctc acatcgcgaa atacaaggct ccatcggtcg
     1261 ggcacatgct tgcccactac cgccgcgacc gcagcagcct ggagcgcgac aacatcgacc
     1321 ccgagcgcat caagaacgac ttggtggtcg gccactacac gaacaaggac ggcaagctcg
     1381 ttgtggggcg cgtggagccg cgcgatggcg agccgaactg gggcacggtg gagagccgca
     1441 tcgagcgcgt gaacgaggcc cagaaggccg cagggaagcg tgccacgcgc aaggatgcgg
     1501 tcgttatggc tgacgttgtt gtgacgcttc ccgacaatgt gcgcaagggc gatgaggaca
     1561 ggttcttccg tctcacgtac tggtacctgt cgaagaagtt cggcatcgag aacatgatgg
     1621 gcgggtacgt ccacaaggac gaggtgttga aggatggcac gcccgcccgc gaccacatgc
     1681 acgtgccgtt cacgcccatc ctagatggac ggttcaacta caagcagatg tgcccccgca
     1741 gcttctaccg gagcatgcac aaggagcttg gggattacct agaggggcgc atgggctacc
     1801 gccctgcaat cgagcttgac gaggagacgc gggcgcagcg tgtatacacg gacaagtccg
     1861 tggatatcga caaggtacgc tgtgcggtcg ataaggccgt ggttgaacct gcgaaggacg
     1921 aagcggcgcg aatcgtcgcg gcggctaggg aagaagccgc cgctttactg aatgatgccg
     1981 aaacgcgtaa ggcagagctt gttaccgaga ttgccgacaa ggagggtgac cttgccgagc
     2041 ttgacagcca gctcgaggac gtgaagctcg atatcgaaga tgagcaagac cgattggagt
     2101 gtcttcggca acgagcggac ggagttgcgc gagacattgg agagcttcaa cccatcgcta
     2161 ccgaggttcg gggctgggaa gctgcgggaa aagctgagcg cggcgcaatc ctcgataaca
     2221 tcgttgttaa gtgcgatgga ctcgcgagcc gcatcagagc aggagttgcg ggaatgcgaa
     2281 tcaaagtgga agagcttaga agtcgaatat cgcgcccgct cgaatatctg atgcgcaggg
     2341 aacagccaag gcaacagagc cttaatgacg tgctgcgcga tgcgcagcgg gcggcggatg
     2401 cgtggaaccg cgaccacgct gcaccgcaga ggacgtatcg cgggcgggct agatgatgtg
     2461 ttatagtagt acctgtaata cacgacaagg aggtgcgacg cgaaaaaacc ccgctgaagc
     2521 ggggtttttt gcgcaccaca gcagacagca gaaaacattc tgaacgctag gaagccggta
     2581 tgattgttac atataagggg aagaaaaatt tcttttaggt acttgctttc ctaaaactga
     2641 tgtgatacaa tgatttaatc cagaaaagga gtaaaaaata tgcggcaagg tattcttaaa
     2701 taaaactata atcaaatagt gggaacaaag gattatgata gctccttttg tagggactta
     2761 gttttttgta cccaatttaa gaatactttt gccttatcaa ttttgacata tccccaaaaa
     2821 cagcaatcac aaacaggtgt atgctgtata tgtgtatgtc cgcaacttat aatccccagt
     2881 ggtaaaagta ttttactgct ggggattttt atgccctttg gggctgtaaa gggaggacaa
     2941 tcacatgaaa ataatcaata ttggaattct tgcccatgta gacgctggaa agacgacctt
     3001 gacggagagc ctgctatatg ccagcggagc catttcagaa ccggggagcg tcgaaaaagg
     3061 gacaacgagg acggacacca tgtttttgga gcggcagcgt gggattacca ttcaagcggc
     3121 agtcacttcc ttccagtggc acagatgtaa agttaacatt gtggatacgc ccggccacat
     3181 ggattttttg gcggaggtgt accgctcttt ggctgtttta gatggggcca tcttggtgat
     3241 ctccgctaaa gatggcgtgc aggcccagac ccgtattctg ttccatgccc tgcggaaaat
     3301 gaacattccc accgttatct ttatcaacaa gatcgaccag gctggcgttg atttgcagag
     3361 cgtggttcag tctgttcggg ataagctctc cgccgatatt atcatcaagc agacggtgtc
     3421 gctgtccccg gaaatagtcc tggaggaaaa taccgacata gaagcatggg atgcggtcat
     3481 cgaaaataac gatgaattat tggaaaagta tatcgcagga gaaccaatca gccgggaaaa
     3541 acttgcgcgg gaggaacagc agcgggttca agacgcctcc ctgttcccag tctatcatgg
     3601 cagcgccaaa aatggccttg gcattcaacc gttgatggat gcggtgacag ggctgttcca
     3661 accgattggg gaacaggggg gcgccgccct atgcggcagc gttttcaagg ttgagtacac
     3721 cgattgcggc cagcggcgtg tctatctacg gttatacagc ggaacgctgc gcctgcggga
     3781 tacggtggcc ctggccggga gagaaaagct gaaaatcaca gagatgcgta ttccatccaa
     3841 aggggaaatt gttcggacag acaccgctta tcagggtgaa attgttatcc ttcccagcga
     3901 cagcgtgagg ttaaacgatg tattagggga ccaaacccgg ctccctcgta aaaggtggcg
     3961 cgaggacccc ctccccatgc tgcggacggc gattgcgccg aaaacggcag cgcaaagaga
     4021 acggctgctg gacgctctta cgcaacttgc ggatactgac ccgcttttgc gttgcgaagt
     4081 ggattccatc acccatgaga tcattctttc ttttttgggc cgggtgcagt tggaggttgt
     4141 ttccgctttg ctgtcggaaa aatacaagct tgaaacagtg gtaaaggaac cctccgtcat
     4201 ttatatggag cggccgctca aagcagccag ccacaccatc catatcgagg tgccgcccaa
     4261 cccgttttgg gcatccatag gactgtctgt tacaccactc tcgcttggct ccggtgtaca
     4321 atacgagagc cgggtttcgc tgggatactt gaaccagagt tttcaaaacg ctgtcaggga
     4381 tggtatccgt tacgggctgg agcagggctt gttcggctgg aacgtaacgg actgtaagat
     4441 ttgctttgaa tacgggcttt attacagtcc ggtcagcacg ccggcggact tccgctcatt
     4501 ggccccgatt gtattggaac aggcattgaa ggaatcgggg acgcagctgc tggaacctta
     4561 tctctccttc atcctctatg cgccccagga atacctttcc agggcttatc atgatgcacc
     4621 gaaatactgt gccaccatcg aaacggccca ggtaaaaaag gatgaagttg tctttactgg
     4681 cgagattccc gcccgctgta tacaggcata ccgtactgat ctggcctttt acaccaacgg
     4741 gcggagcgta tgccttacag agctgaaagg atatcaggcc gctgtcggtc agccggtcat
     4801 ccagccccgc cgtccaaaca gccgcctgga caaggtgcgc catatgtttc agaaggtaat
     4861 gtaaatcgtt aagggatcaa ctttgggaga gagttcaaaa ttgatccttt ttttataaca
     4921 ggaattaaat aataaaaaag ccggattaat aatctggctt tttatattct ctctctagta
     4981 tataaacgca gaaaggccca cccgaaggtg agccagtgtg agtcgactag ttcacgcgca
     5041 cggatccacc gggtcggccg ggtcgtcggc ggcctcttcg ggcaccgtgc ccatcaggct
     5101 gacctcgccg atgccgattt tgatgaggtc ggtctccgtg aacaggaaca aatgcgcgat
     5161 gaacagcagc gacacggcca ccagcgtgac cagcgtgagg tcgaacatgc tgaccccggt
     5221 cgcctccagc gaatgggcga acagctggcc tgcggtctcg ccgccgtaca gcgccacgaa
     5281 ccccaggccg atcgtccgcg ccgggctcac gcggaagctg gacgccacgt cgatgcacac
     5341 cacgcacagc accagcacga agcagtggta gccgccgaag ataagcgcgc tcgaatacgt
     5401 gttgttgtcg gccatgaacg gcgtggacaa gcatcccaag cacagcatca gcatggagac
     5461 gcggtacgag aagaacacgc tgtcgcgctg caggaacacg gctatcaggc agatggccaa
     5521 cgctccgatg aagcccgcca gaaacaggta cgtgccgaag atgtcctgca cctcgtaggt
     5581 tttgaacccc gacaccacgt cgaagaaccc catcaacatg ccgatgagca ccgcgcccag
     5641 aagcgccttc gcgaacaggg tcgccgtatg gcgcgacaaa ggctgctcgg gctcgcgcgg
     5701 cagctgcggc gtatcgggcg cgctgcgcct gagagctgcc gccgaaagca cgggcatgac
     5761 aacgatgagc acgatgacgg gtatcgccgg ccctacgatc agcaagaacc ccacgacgaa
     5821 cgcgatgcac agcgacgagg caagctccac aagcgccaag cggcctccca gccccgactc
     5881 gagctcgatc cagcacaaga acagcacggc cgagcccgtg ccggtcatga ccgcgctcag
     5941 gcccagcacc agcatgccga cgggcgtggt aggatcggag aacggcgtga gcacgctgcc
     6001 cgcgctcatc atgatggccg atcccagcac gatggagcgg ttgccgctca cccgctcgcg
     6061 ccgcacgatg acggagacga acagcatggt caccaccatg gccaccagcg atatgatcag
     6121 gctgacgtac agcgagccgc gcacgtccga cgagaagccg tacggcacga tgctgtagaa
     6181 catcatgagc atccaaagcc attgcaaggc cgcgcccgcg atgcgcttga gggaaaggct
     6241 cgtccccaac gcggtatccc atgcactccg aagcttttcg gatccgcggc cgctacgtag
     6301 aattcccttg tcatcgtcat ccttgtaatc gatgtcatga tctttataat caccgtcatg
     6361 gtctttgtag tccatgcccc accccctgtg agcccgtaca taacgtcgct cgcgccgacc
     6421 cctctgtcga cgggttcgat tatacggaaa agatgagcgg ccctctttga ggaaactatt
     6481 ctagtttcag aataatttat gacctacggt taaagtttcc ccagttcaaa gcactatcga
     6541 atttttcatg atcacaagat cataagaatt tatgatgcgg tcgaacctgc tttccaacct
     6601 ttggacgatg ctcatctaga agcggccgcg aattcagaat agtaatacag gatccgaatc
     6661 gtttcagttg ccgccggatt aataatcaaa taaccgttcc taaaaaatgc cctcttgggt
     6721 tatcaagagg gtcattatat ttcgcaaaaa accccgcttc ggcggggttt tttcgccttg
     6781 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6841 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7201 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     7261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7501 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     7561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7861 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7921 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7981 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8101 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8161 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8221 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8341 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8401 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8461 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8521 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8581 aggggggcgg agcctatgga aaaacgccag caacgcggcc cgagatgcgc cgcgtgcggc
     8641 tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag
     8701 ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc
     8761 gccggcatga gctcctctag tagagagcgt tcaccgacaa acaacagata aaacgaaagg
     8821 cccagtcttt cgactgagcc tttcgtttta tttgaaaaag aaaagcccca gcaagcttgg
     8881 cggcagcagg ggca
//
