LOCUS       sequence-247783-        4678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-159aa)-ydfI(153-213aa) chimera under PLtetO-1, 
            output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4678)
  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 4678)
  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: pSR357.24
FEATURES             Location/Qualifiers
     source          1..4678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..81
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(102..120,121..126,127..143,144..149,150..175)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    102..120
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    127..145
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      893..964
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      980..1007
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1246..1257
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1264..1977
                     /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      2004..2075
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2091..2118
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2170..2204
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2239..2865
                     /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
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2315..2334)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2871..2957
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3122..3710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3202..3221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3798..3892)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3913..4572)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4487..4506
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tctcattttc gccagatatc gacgtctaag aaaccattat tatcatgaca ttaacctata
       61 aaaataggcg tatcacgagg ccctttcgtc ttcacctcga gtccctatca gtgatagaga
      121 ttgacatccc tatcagtgat agagatactg agcacatcag caggacgcac tgacctaagg
      181 attttttcta gaagaattat atacaacaaa tgagtaatca ggaaccggct actatcctgc
      241 tgattgacga tcacccgatg ctgcgaactg gcgtaaaaca gcttatcagt atggcaccag
      301 atatcaccgt ggttggcgaa gcgagtaatg gcgaacaggg tattgaactg gcggagtctc
      361 ttgatcccga tctgatcctg ttagatctca atatgcccgg catgaacggt ctggaaacgc
      421 tggataaact gcgcgaaaag tccctctcag ggcgcattgt ggtattcagc gtctctaacc
      481 atgaagaaga tgtggtcacc gcactgaaac gcggcgcgga tggctatctg ttaaaagata
      541 tggaaccgga agatctgctg aaagcattgc atcaggcagc tgctggcgaa atggtattaa
      601 gcgaagcatt aacgcctgtt ctggccgcca gcttgcgcgc taaccgtgcc actactgagc
      661 gcgatgttaa ccagttaacc ccacgcgaag tcattgttct aaaagcaatt gctaaaggtc
      721 ttaaaagcaa agcgattgcc tttgatttgg gcgtctctga gcgaacagta aagtccagat
      781 taacgtccat ttacaataaa ttaggcgcga attcaagaac tgaggcagta acgattgcca
      841 tgcaaaaagg tattctgaca atagacaact aataataatc tagaccaggc atcaaataaa
      901 acgaaaggat cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      961 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atagtaaatc
     1021 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcaccga
     1081 catcataacg gttctggcaa atattctgaa atgagctggt tagctagtca actgcatatt
     1141 tgaaaattgc ccaaacgtac atgcccgaat gtacgttttt ttcatttcat tgtcaactac
     1201 aatgagaaag aatgtgatca agcaatgtgt tgaaaggaga ttatcaaaga ggagaaatac
     1261 tagatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat
     1321 ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat
     1381 ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc ttggccgact
     1441 ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag
     1501 cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc
     1561 tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg
     1621 gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat
     1681 aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat
     1741 ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct
     1801 gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac
     1861 tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt
     1921 ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaatga
     1981 tgataataat ctagaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     2041 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     2101 cgggtgggcc tttctgcgtt tataggtacc cgtggatcct ctagagtcga cctgcaggca
     2161 tgcaagcttt ttacggctag ctcagtccta ggtatagtgc tagcccagcc agagaaacac
     2221 tctttaacag ggggtagtat gatgtctcgt ttagataaaa gtaaagtgat taacagcgca
     2281 ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag
     2341 ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac
     2401 gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa
     2461 agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat
     2521 cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc
     2581 gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca
     2641 ctcagcgcag tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc
     2701 gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct
     2761 atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc
     2821 atatgcggat tagaaaaaca acttaaatgt gaaagtgggt cttaaggcat caaataaaac
     2881 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2941 tcctgagtag gacaaatccg ccgccctaga cctatggcgt tcggctgcgg cgagcggtat
     3001 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcatgcaaga
     3061 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3121 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3181 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3241 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3301 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3361 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3421 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3481 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3541 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3601 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3661 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3721 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3781 tcatgactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     3841 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
     3901 ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     3961 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     4021 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     4081 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     4141 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cataacacgc
     4201 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     4261 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     4321 tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg
     4381 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     4441 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     4501 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     4561 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgac
     4621 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgggcc ggccaacg
//
