LOCUS       sequence-375874-        9110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli - Eggerthella lenta shuttle plasmid (TetR), 
            N-3xFLAG-dadR(linker region shuffled) 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(DadRmt7)-N-3xFLAG-DadR(linker)
FEATURES             Location/Qualifiers
     source          1..9110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(4972..4999)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6508..6573)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    6817..6836
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     primer_bind     complement(6993..7011)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7079..7183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7184..7252,7253..8044)
                     /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(7402..7421)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8215..8803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8704..8723
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(8984..9055)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttatagcatg gcggcaaacg ctggcgagac gaaggtgaag ctctgggagg gcatctgcta
       61 cccagagaac atgcgcgatg actggcagga cgagatagac gacattcttc aagtgccttt
      121 cgcttatgcg gttcacgaca tagaccatga ccagaagagc aagcagcgca agacgcatgc
      181 gcacgtcatt gttgtctggg gcggcaacac gacacgcaag gcaatcatca acgtcttgaa
      241 ccgattgagc gcggacggca agaagtgctg ttcgtcagcc gagccaatca acaacatcag
      301 gcatgcgtat gactatctga ttcacgacac ggcttcctgt cgcaagaagg gcaaggagct
      361 gtatccggtc gaagcacgca tcgagggaaa caacttcgac attggtcagc tcgaacagct
      421 ctcgactaag gacaagcagg acatgctctt cgagttggtc ggcttcatca tgtgcgagag
      481 gttcgagacc atcaacgatt tcacggctgc ggcgctgagg gagtttcccg agcagtaccg
      541 cgaaatcatc gtggggtaca actcgattct cgagcgctat tgtcgcggga actatctgaa
      601 cgccgaacgt aagcgcaagt gcgtggaaca gtcaggtgcc aaagagtagt cgggtcaatg
      661 cagcaatgcg tgactacggg gcggcgtggt ctgcgctcgc gcagatctaa gccgactcgt
      721 tcacggattg cggaattgac gcgacggcca accgaataga ccagacttga ttaagggtcg
      781 atggacgctc tgctccatcg accctttcct tgcccgtgtc tcgactggat gagacctgca
      841 gaatcgggcg gctcattatt tttcgctaac tacgtccgaa aaataatgtg tccgaaaaat
      901 aatgtgacaa atgacaaatg acaaatgacc acaggggggc cactacgacg gcccccctaa
      961 tagtgccgtg tgccgtgtgc cgaaccgcag gtagattgag gggtggttcg gaggggcgaa
     1021 gtccctccgt tcggcgctcg cgccgatggc ccgagggccg tgagcgaagc cgcgaacggg
     1081 caagggctgg ggggtctcgg ggggaacgcc cgagcgggtg cagggctggc cctgccacgt
     1141 ccttagcgta gctaaggtct aacgtgctag actttagtcg attaagtctg tacggttcgg
     1201 aggtttccat ggctcacatc gcgaaataca aggctccatc ggtcgggcac atgcttgccc
     1261 actaccgccg cgaccgcagc agcctggagc gcgacaacat cgaccccgag cgcatcaaga
     1321 acgacttggt ggtcggccac tacacgaaca aggacggcaa gctcgttgtg gggcgcgtgg
     1381 agccgcgcga tggcgagccg aactggggca cggtggagag ccgcatcgag cgcgtgaacg
     1441 aggcccagaa ggccgcaggg aagcgtgcca cgcgcaagga tgcggtcgtt atggctgacg
     1501 ttgttgtgac gcttcccgac aatgtgcgca agggcgatga ggacaggttc ttccgtctca
     1561 cgtactggta cctgtcgaag aagttcggca tcgagaacat gatgggcggg tacgtccaca
     1621 aggacgaggt gttgaaggat ggcacgcccg cccgcgacca catgcacgtg ccgttcacgc
     1681 ccatcctaga tggacggttc aactacaagc agatgtgccc ccgcagcttc taccggagca
     1741 tgcacaagga gcttggggat tacctagagg ggcgcatggg ctaccgccct gcaatcgagc
     1801 ttgacgagga gacgcgggcg cagcgtgtat acacggacaa gtccgtggat atcgacaagg
     1861 tacgctgtgc ggtcgataag gccgtggttg aacctgcgaa ggacgaagcg gcgcgaatcg
     1921 tcgcggcggc tagggaagaa gccgccgctt tactgaatga tgccgaaacg cgtaaggcag
     1981 agcttgttac cgagattgcc gacaaggagg gtgaccttgc cgagcttgac agccagctcg
     2041 aggacgtgaa gctcgatatc gaagatgagc aagaccgatt ggagtgtctt cggcaacgag
     2101 cggacggagt tgcgcgagac attggagagc ttcaacccat cgctaccgag gttcggggct
     2161 gggaagctgc gggaaaagct gagcgcggcg caatcctcga taacatcgtt gttaagtgcg
     2221 atggactcgc gagccgcatc agagcaggag ttgcgggaat gcgaatcaaa gtggaagagc
     2281 ttagaagtcg aatatcgcgc ccgctcgaat atctgatgcg cagggaacag ccaaggcaac
     2341 agagccttaa tgacgtgctg cgcgatgcgc agcgggcggc ggatgcgtgg aaccgcgacc
     2401 acgctgcacc gcagaggacg tatcgcgggc gggctagatg atgtgttata gtagtacctg
     2461 taatacacga caaggaggtg cgacgcgaaa aaaccccgct gaagcggggt tttttgcgca
     2521 ccacagcaga cagcagaaaa cattctgaac gctaggaagc cggtatgatt gttacatata
     2581 aggggaagaa aaatttcttt taggtacttg ctttcctaaa actgatgtga tacaatgatt
     2641 taatccagaa aaggagtaaa aaatatgcgg caaggtattc ttaaataaaa ctataatcaa
     2701 atagtgggaa caaaggatta tgatagctcc ttttgtaggg acttagtttt ttgtacccaa
     2761 tttaagaata cttttgcctt atcaattttg acatatcccc aaaaacagca atcacaaaca
     2821 ggtgtatgct gtatatgtgt atgtccgcaa cttataatcc ccagtggtaa aagtatttta
     2881 ctgctgggga tttttatgcc ctttggggct gtaaagggag gacaatcaca tgaaaataat
     2941 caatattgga attcttgccc atgtagacgc tggaaagacg accttgacgg agagcctgct
     3001 atatgccagc ggagccattt cagaaccggg gagcgtcgaa aaagggacaa cgaggacgga
     3061 caccatgttt ttggagcggc agcgtgggat taccattcaa gcggcagtca cttccttcca
     3121 gtggcacaga tgtaaagtta acattgtgga tacgcccggc cacatggatt ttttggcgga
     3181 ggtgtaccgc tctttggctg ttttagatgg ggccatcttg gtgatctccg ctaaagatgg
     3241 cgtgcaggcc cagacccgta ttctgttcca tgccctgcgg aaaatgaaca ttcccaccgt
     3301 tatctttatc aacaagatcg accaggctgg cgttgatttg cagagcgtgg ttcagtctgt
     3361 tcgggataag ctctccgccg atattatcat caagcagacg gtgtcgctgt ccccggaaat
     3421 agtcctggag gaaaataccg acatagaagc atgggatgcg gtcatcgaaa ataacgatga
     3481 attattggaa aagtatatcg caggagaacc aatcagccgg gaaaaacttg cgcgggagga
     3541 acagcagcgg gttcaagacg cctccctgtt cccagtctat catggcagcg ccaaaaatgg
     3601 ccttggcatt caaccgttga tggatgcggt gacagggctg ttccaaccga ttggggaaca
     3661 ggggggcgcc gccctatgcg gcagcgtttt caaggttgag tacaccgatt gcggccagcg
     3721 gcgtgtctat ctacggttat acagcggaac gctgcgcctg cgggatacgg tggccctggc
     3781 cgggagagaa aagctgaaaa tcacagagat gcgtattcca tccaaagggg aaattgttcg
     3841 gacagacacc gcttatcagg gtgaaattgt tatccttccc agcgacagcg tgaggttaaa
     3901 cgatgtatta ggggaccaaa cccggctccc tcgtaaaagg tggcgcgagg accccctccc
     3961 catgctgcgg acggcgattg cgccgaaaac ggcagcgcaa agagaacggc tgctggacgc
     4021 tcttacgcaa cttgcggata ctgacccgct tttgcgttgc gaagtggatt ccatcaccca
     4081 tgagatcatt ctttcttttt tgggccgggt gcagttggag gttgtttccg ctttgctgtc
     4141 ggaaaaatac aagcttgaaa cagtggtaaa ggaaccctcc gtcatttata tggagcggcc
     4201 gctcaaagca gccagccaca ccatccatat cgaggtgccg cccaacccgt tttgggcatc
     4261 cataggactg tctgttacac cactctcgct tggctccggt gtacaatacg agagccgggt
     4321 ttcgctggga tacttgaacc agagttttca aaacgctgtc agggatggta tccgttacgg
     4381 gctggagcag ggcttgttcg gctggaacgt aacggactgt aagatttgct ttgaatacgg
     4441 gctttattac agtccggtca gcacgccggc ggacttccgc tcattggccc cgattgtatt
     4501 ggaacaggca ttgaaggaat cggggacgca gctgctggaa ccttatctct ccttcatcct
     4561 ctatgcgccc caggaatacc tttccagggc ttatcatgat gcaccgaaat actgtgccac
     4621 catcgaaacg gcccaggtaa aaaaggatga agttgtcttt actggcgaga ttcccgcccg
     4681 ctgtatacag gcataccgta ctgatctggc cttttacacc aacgggcgga gcgtatgcct
     4741 tacagagctg aaaggatatc aggccgctgt cggtcagccg gtcatccagc cccgccgtcc
     4801 aaacagccgc ctggacaagg tgcgccatat gtttcagaag gtaatgtaaa tcgttaaggg
     4861 atcaactttg ggagagagtt caaaattgat ccttttttta taacaggaat taaataataa
     4921 aaaagccgga ttaataatct ggctttttat attctctctc tagtatataa acgcagaaag
     4981 gcccacccga aggtgagcca gtgtgagtcg actagttcag accgcctcgt cctgcgcggc
     5041 cgtctcgtgg gccaggtcga tgagctcctg gcgattgtcc acgcccgtct tctggtagat
     5101 gtggcggata tgcgtgctca ccgtgcccgc cgagatgaac agcgtctcct ggatgcgcga
     5161 gatggtgcgg ccctccagca gcagcggaag cacgtcggtc tcgcgcggcg acagggcgaa
     5221 gcgctccgcg atcagcgcgc acggggcgtc cgggtcgtcg cccgtggggg gttcggcggc
     5281 cacctccaca tccatcaggc tgacctcgcc gatgccgatt ttgatgaggt cggtctccgt
     5341 gaacaggaac aaatgcgcga tgaacagcag cgacacggcc accagcgtga ccagcgtgag
     5401 gtcgaacatg ctgaccccgg tcgcctccag cgaatgggcg aacagctggc ctgcggtctc
     5461 gccgccgtac agcgccacga accccaggcc gatcgtccgc gccgggctca cgcggaagct
     5521 ggacgccacg tcgatgcaca ccacgcacag caccagcacg aagcagtggt agccgccgaa
     5581 gataagcgcg ctcgaatacg tgttgttgtc ggccatgaac ggcgtggaca agcatcccaa
     5641 gcacagcatc agcatggaga cgcggtacga gaagaacacg ctgtcgcgct gcaggaacac
     5701 ggctatcagg cagatggcca acgctccgat gaagcccgcc agaaacaggt acgtgccgaa
     5761 gatgtcctgc acctcgtagg ttttgaaccc cgacaccacg tcgaagaacc ccatcaacat
     5821 gccgatgagc accgcgccca gaagcgcctt cgcgaacagg gtcgccgtat ggcgcgacaa
     5881 aggctgctcg ggctcgcgcg gcagctgcgg cgtatcgggc gcgctgcgcc tgagagctgc
     5941 cgccgaaagc acgggcatga caacgatgag cacgatgacg ggtatcgccg gccctacgat
     6001 cagcaagaac cccacgacga acgcgatgca cagcgacgag gcaagctcca caagcgccaa
     6061 gcggcctccc agccccgact cgagctcgat ccagcacaag aacagcacgg ccgagcccgt
     6121 gccggtcatg accgcgctca ggcccagcac cagcatgccg acgggcgtgg taggatcgga
     6181 gaacggcgtg agcacgctgc ccgcgctcat catgatggcc gatcccagca cgatggagcg
     6241 gttgccgctc acccgctcgc gccgcacgat gacggagacg aacagcatgg tcaccaccat
     6301 ggccaccagc gatatgatca ggctgacgta cagcgagccg cgcacgtccg acgagaagcc
     6361 gtacggcacg atgctgtaga acatcatgag catccaaagc cattgcaagg ccgcgcccgc
     6421 gatgcgcttg agggaaaggc tcgtccccaa cgcggtatcc catgcactcc gaagcttttc
     6481 ggatccgcgg ccgctacgta gaattccctt gtcatcgtca tccttgtaat cgatgtcatg
     6541 atctttataa tcaccgtcat ggtctttgta gtccatgccc caccccctgt gagcccgtac
     6601 ataacgtcgc tcgcgccgac ccctctgtcg acgggttcga ttatacggaa aagatgagcg
     6661 gccctctttg aggaaactat tctagtttca gaataattta tgacctacgg ttaaagtttc
     6721 cccagttcaa agcactatcg aatttttcat gatcacaaga tcataagaat ttatgatgcg
     6781 gtcgaacctg ctttccaacc tttggacgat gctcatctag aagcggccgc gaattcagaa
     6841 tagtaataca ggatccgaat cgtttcagtt gccgccggat taataatcaa ataaccgttc
     6901 ctaaaaaatg ccctcttggg ttatcaagag ggtcattata tttcgcaaaa aaccccgctt
     6961 cggcggggtt ttttcgcctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     7021 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7081 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7141 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7201 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7261 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7321 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7381 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     7441 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7501 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7561 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7621 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7681 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     7741 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7801 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7861 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7921 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7981 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8041 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8101 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8161 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8221 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8281 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8341 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     8401 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8461 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8521 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8581 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8641 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8701 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8761 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8821 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     8881 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     8941 tgaatccgtt agcgaggtgc cgccggcatg agctcctcta gtagagagcg ttcaccgaca
     9001 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgaaaaa
     9061 gaaaagcccc agcaagcttg gcggcagcag gggcattcga gacaaggaga
//
