LOCUS       sequence-375884-        9110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli - Eggerthella lenta shuttle plasmid (TetR), N-3xFLAG-dadR 
            complementation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9110)
  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 9110)
  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(DadR-NFLAG)-N-3xFLAG-DadR
FEATURES             Location/Qualifiers
     source          1..9110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(4987..5014)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6523..6588)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    6832..6851
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     primer_bind     complement(7008..7026)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7094..7198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7199..7267,7268..8059)
                     /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(7417..7436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8230..8818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8719..8738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(8999..9070)
                     /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 ttcagaccgc
     5041 ctcgtcctgc gcggccgtct cgtgggccag gtcgatgagc tcctggcgat tgtccacgcc
     5101 cgtcttctgg tagatgtggc ggatatgcgt gctcaccgtg cccgccgaga tgaacagcgt
     5161 ctcctggatg cgcgagatgg tgcggccctc cagcagcagc ggaagcacgt cggtctcgcg
     5221 cggcgacagg gcgaagcgct ccgcgatcag cgcgcacgga tccaccgggt cggccgggtc
     5281 gtcggcggcc tcttcgggca ccgtgcccat caggctgacc tcgccgatgc cgattttgat
     5341 gaggtcggtc tccgtgaaca ggaacaaatg cgcgatgaac agcagcgaca cggccaccag
     5401 cgtgaccagc gtgaggtcga acatgctgac cccggtcgcc tccagcgaat gggcgaacag
     5461 ctggcctgcg gtctcgccgc cgtacagcgc cacgaacccc aggccgatcg tccgcgccgg
     5521 gctcacgcgg aagctggacg ccacgtcgat gcacaccacg cacagcacca gcacgaagca
     5581 gtggtagccg ccgaagataa gcgcgctcga atacgtgttg ttgtcggcca tgaacggcgt
     5641 ggacaagcat cccaagcaca gcatcagcat ggagacgcgg tacgagaaga acacgctgtc
     5701 gcgctgcagg aacacggcta tcaggcagat ggccaacgct ccgatgaagc ccgccagaaa
     5761 caggtacgtg ccgaagatgt cctgcacctc gtaggttttg aaccccgaca ccacgtcgaa
     5821 gaaccccatc aacatgccga tgagcaccgc gcccagaagc gccttcgcga acagggtcgc
     5881 cgtatggcgc gacaaaggct gctcgggctc gcgcggcagc tgcggcgtat cgggcgcgct
     5941 gcgcctgaga gctgccgccg aaagcacggg catgacaacg atgagcacga tgacgggtat
     6001 cgccggccct acgatcagca agaaccccac gacgaacgcg atgcacagcg acgaggcaag
     6061 ctccacaagc gccaagcggc ctcccagccc cgactcgagc tcgatccagc acaagaacag
     6121 cacggccgag cccgtgccgg tcatgaccgc gctcaggccc agcaccagca tgccgacggg
     6181 cgtggtagga tcggagaacg gcgtgagcac gctgcccgcg ctcatcatga tggccgatcc
     6241 cagcacgatg gagcggttgc cgctcacccg ctcgcgccgc acgatgacgg agacgaacag
     6301 catggtcacc accatggcca ccagcgatat gatcaggctg acgtacagcg agccgcgcac
     6361 gtccgacgag aagccgtacg gcacgatgct gtagaacatc atgagcatcc aaagccattg
     6421 caaggccgcg cccgcgatgc gcttgaggga aaggctcgtc cccaacgcgg tatcccatgc
     6481 actccgaagc ttttcggatc cgcggccgct acgtagaatt cccttgtcat cgtcatcctt
     6541 gtaatcgatg tcatgatctt tataatcacc gtcatggtct ttgtagtcca tgccccaccc
     6601 cctgtgagcc cgtacataac gtcgctcgcg ccgacccctc tgtcgacggg ttcgattata
     6661 cggaaaagat gagcggccct ctttgaggaa actattctag tttcagaata atttatgacc
     6721 tacggttaaa gtttccccag ttcaaagcac tatcgaattt ttcatgatca caagatcata
     6781 agaatttatg atgcggtcga acctgctttc caacctttgg acgatgctca tctagaagcg
     6841 gccgcgaatt cagaatagta atacaggatc cgaatcgttt cagttgccgc cggattaata
     6901 atcaaataac cgttcctaaa aaatgccctc ttgggttatc aagagggtca ttatatttcg
     6961 caaaaaaccc cgcttcggcg gggttttttc gccttgaaga cgaaagggcc tcgtgatacg
     7021 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     7081 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7141 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     7201 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     7261 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     7321 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     7381 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     7441 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     7501 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7561 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7621 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7681 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7741 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7801 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7861 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7921 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7981 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     8041 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     8101 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8161 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8221 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     8281 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     8341 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     8401 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     8461 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     8521 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8581 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8641 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8701 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8761 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8821 cgccagcaac gcggcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg
     8881 atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc
     8941 caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcatgagctc ctctagtaga
     9001 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     9061 gttttatttg aaaaagaaaa gccccagcaa gcttggcggc agcaggggca
//
