LOCUS       sequence-247753-        4675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-126aa)-ydfI(121-213aa) chimera under PLtetO-1, 
            output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4675)
  AUTHORS   Schmidl SR, Ekness F, Sofjan K, Daeffler KN, Brink KR, Landry BP, 
            Gerhardt KP, Dyulgyarov N, Sheth RU, Tabor JJ
  TITLE     Rewiring bacterial two-component systems by modular DNA-binding 
            domain swapping.
  JOURNAL   Nat Chem Biol. 2019 Jul;15(7):690-698. doi: 
            10.1038/s41589-019-0286-6. Epub 2019 May 20.
  PUBMED    31110305
REFERENCE   2  (bases 1 to 4675)
  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"; doi: 
            "10.1038/s41589-019-0286-6"; journalName: "Nat Chem Biol"; date: 
            "2019-07"; volume: "15"; issue: "7"; pages: "690-698"
COMMENT     Sequence Label: pSR357.6
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..24
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(45..63,64..69,70..86,87..92,93..118)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    70..88
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      833..904
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      920..947
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1186..1197
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1204..1917
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1944..2015
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2031..2058
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2110..2144
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2179..2805
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESWS"
     primer_bind     complement(2255..2274)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2811..2897
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3062..3650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3142..3161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3738..3832)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3853..4512)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4427..4446
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ataaaaatag gcgtatcacg aggccctttc gtcttcacct cgagtcccta tcagtgatag
       61 agattgacat ccctatcagt gatagagata ctgagcacat cagcaggacg cactgaccta
      121 aggatttttt ctagaagaat tatatacaac aaatgagtaa tcaggaaccg gctactatcc
      181 tgctgattga cgatcacccg atgctgcgaa ctggcgtaaa acagcttatc agtatggcac
      241 cagatatcac cgtggttggc gaagcgagta atggcgaaca gggtattgaa ctggcggagt
      301 ctcttgatcc cgatctgatc ctgttagatc tcaatatgcc cggcatgaac ggtctggaaa
      361 cgctggataa actgcgcgaa aagtccctct cagggcgcat tgtggtattc agcgtctcta
      421 accatgaaga agatgtggtc accgcactga aacgcggcgc ggatggctat ctgttaaaag
      481 atatggaacc ggaagatctg ctgaaagcat tgcatcaggc agctgctggc aatgtgctat
      541 tgcagcctga tatccttaaa cgtctgcaag aaatccaatt tgagcggatg aaaaagcagc
      601 gcaatgagac gcagcttaca gaaaaggaag tcattgttct aaaagcaatt gctaaaggtc
      661 ttaaaagcaa agcgattgcc tttgatttgg gcgtctctga gcgaacagta aagtccagat
      721 taacgtccat ttacaataaa ttaggcgcga attcaagaac tgaggcagta acgattgcca
      781 tgcaaaaagg tattctgaca atagacaact aataataatc tagaccaggc atcaaataaa
      841 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      901 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atagtaaatc
      961 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga
     1021 catcataacg gttctggcaa atattctgaa atgagctggt tagctagtca actgcatatt
     1081 tgaaaattgc ccaaacgtac atgcccgaat gtacgttttt ttcatttcat tgtcaactac
     1141 aatgataaag aatgtgatca agcaatgtgt tgaaaggaga ttatcaaaga ggagaaatac
     1201 tagatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat
     1261 ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat
     1321 ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc ttggccgact
     1381 ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag
     1441 cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc
     1501 tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg
     1561 gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat
     1621 aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat
     1681 ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct
     1741 gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac
     1801 tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt
     1861 ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaatga
     1921 tgataataat ctagaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1981 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     2041 cgggtgggcc tttctgcgtt tataggtacc cgtggatcct ctagagtcga cctgcaggca
     2101 tgcaagcttt ttacggctag ctcagtccta ggtatagtgc tagcccagcc agagaaacac
     2161 tctttaacag ggggtagtat gatgtctcgt ttagataaaa gtaaagtgat taacagcgca
     2221 ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag
     2281 ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac
     2341 gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa
     2401 agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat
     2461 cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc
     2521 gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca
     2581 ctcagcgcag tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc
     2641 gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct
     2701 atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc
     2761 atatgcggat tagaaaaaca acttaaatgt gaaagttggt cttaaggcat caaataaaac
     2821 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2881 tcctgagtag gacaaatccg ccgccctaga cctagggcgt tcggctgcgg cgagcggtat
     2941 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcatgcaaga
     3001 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3481 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3721 tcatgactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     3781 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
     3841 ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     3901 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     3961 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     4021 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     4081 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cataacacgc
     4141 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     4201 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     4261 tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg
     4321 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     4381 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     4441 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     4501 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgac
     4561 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgggcc ggccaacgtc
     4621 tcattttcgc cagatatcga cgtctaagaa accattatta tcatgacatt aacct
//
