LOCUS       sequence-334946-        4625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Feedback-controlled AraC circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4625)
  AUTHORS   Bongaerts N, Edoo Z, Abukar AA, Song X, Sosa-Carrillo S, 
            Haggenmueller S, Savigny J, Gontier S, Lindner AB, Wintermute EH
  TITLE     Low-cost anti-mycobacterial drug discovery using engineered E. coli.
  JOURNAL   Nat Commun. 2022 Jul 7;13(1):3905. doi: 10.1038/s41467-022-31570-3.
  PUBMED    35798732
REFERENCE   2  (bases 1 to 4625)
  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: "2022-07-7"; volume: "13(1):3905. doi"; pages: " 
            10.1038/s41467-022-31570-3"
COMMENT     Sequence Label: pNB101
FEATURES             Location/Qualifiers
     source          1..4625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..291
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     214..233
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             302..313
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             320..1198
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      1234..1320
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1858..2808)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2856..3078
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3570..3664)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3695..4489)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3806..3825)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4416..4435
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact
       61 ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg
      121 ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc
      181 acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat
      241 tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca tagaattcat
      301 taaagaggag aaaggtacca tggctgaagc gcaaaatgat cccctgctgc cgggatactc
      361 gtttaacgcc catctggtgg cgggtttaac gccgattgag gccaacggtt atctcgattt
      421 ttttatcgac cgaccgctgg gaatgaaagg ttatattctc aatctcacca ttcgcggtca
      481 gggggtggtg aaaaatcagg gacgagaatt tgtctgccga ccgggtgata ttttgctgtt
      541 cccgccagga gagattcatc actacggtcg tcatccggag gctcgcgaat ggtatcacca
      601 gtgggtttac tttcgtccgc gcgcctactg gcatgaatgg cttaactggc cgtcaatatt
      661 tgccaatacg ggtttctttc gcccggatga agcgcaccag ccgcatttca gcgacctgtt
      721 tgggcaaatc attaacgccg ggcaagggga agggcgctat tcggagctgc tggcgataaa
      781 tctgcttgag caattgttac tgcggcgcat ggaagcgatt aacgagtcgc tccatccacc
      841 gatggataat cgggtacgcg aggcttgtca gtacatcagc gatcacctgg cagacagcaa
      901 ttttgatatc gccagcgtcg cacagcatgt ttgcttgtcg ccgtcgcgtc tgtcacatct
      961 tttccgccag cagttaggga ttagcgtctt aagctggcgc gaggaccaac gcattagtca
     1021 ggcgaagctg cttttgagca ctacccggat gcctatcgcc accgtcggtc gcaatgttgg
     1081 ttttgacgat caactctatt tctcgcgagt atttaaaaaa tgcaccgggg ccagcccgag
     1141 cgagtttcgt gccggttgtg aagaaaaagt gaatgatgta gccgtcaagt tgtcataacc
     1201 cgggggatcc catggtacgc gtgctagagg catcaaataa aacgaaaggc tcagtcgaaa
     1261 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1321 ccgccgccct agacctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg
     1381 ctagtttgtt atcagaatcg cagatccggc ttcagccggt ttgccggctg aaagcgctat
     1441 ttcttccaga attgccatga ttttttcccc acgggaggcg tcactggctc ccgtgttgtc
     1501 ggcagctttg attcgataag cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga
     1561 gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt
     1621 tcacctgttc tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg
     1681 tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca
     1741 ccgttttcat ctgtgcatat ggacagtttt ccctttgata tgtaacggtg aacagttgtt
     1801 ctacttttgt ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca
     1861 gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca
     1921 tgagaacgaa ccattgagat catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa
     1981 actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttatg
     2041 taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt
     2101 gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt
     2161 atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc
     2221 tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc
     2281 aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt
     2341 cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa
     2401 agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa
     2461 tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca
     2521 ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag
     2581 ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg
     2641 agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt
     2701 ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata
     2761 gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg
     2821 gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt
     2881 ccttttcctt tgagttgtgg gtatctgtaa attctgctag acctttgctg gaaaacttgt
     2941 aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat
     3001 tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc
     3061 cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca
     3121 aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct
     3181 cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc
     3241 gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta
     3301 aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa
     3361 agcccgtcac gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac
     3421 tttttgctgt tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc
     3481 cgtgacaggt cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacaggc
     3541 ttacccgtct tactgtccct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     3601 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     3661 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     3721 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     3781 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     3841 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     3901 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     3961 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     4021 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     4081 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     4141 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     4201 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     4261 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     4321 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     4381 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     4441 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     4501 ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     4561 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     4621 attcc
//
