LOCUS       sequence-247787-        4675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-122aa)-ydfI(117-213aa) 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: pSR357.28
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..145
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(166..184,185..190,191..207,208..213,214..239)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    166..184
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    191..209
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      954..1025
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1041..1068
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1307..1318
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1325..2038
                     /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      2065..2136
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2152..2179
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2231..2265
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2300..2926
                     /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(2376..2395)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2932..3018
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3183..3771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3263..3282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3859..3953)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3974..4633)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4548..4567
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tacgaccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgggccggcc
       61 aacgtctcat tttcgccaga tatcgacgtc taagaaacca ttattatcat gacattaacc
      121 tataaaaata ggcgtatcac gaggcccttt cgtcttcacc tcgagtccct atcagtgata
      181 gagattgaca tccctatcag tgatagagat actgagcaca tcagcaggac gcactgacct
      241 aaggattttt tctagaagaa ttatatacaa caaatgagta atcaggaacc ggctactatc
      301 ctgctgattg acgatcaccc gatgctgcga actggcgtaa aacagcttat cagtatggca
      361 ccagatatca ccgtggttgg cgaagcgagt aatggcgaac agggtattga actggcggag
      421 tctcttgatc ccgatctgat cctgttagat ctcaatatgc ccggcatgaa cggtctggaa
      481 acgctggata aactgcgcga aaagtccctc tcagggcgca ttgtggtatt cagcgtctct
      541 aaccatgaag aagatgtggt caccgcactg aaacgcggcg cggatggcta tctgttaaaa
      601 gatatggaac cggaagatct gctgaaagca ttgcatcagg ccatcagagg aaatgtgcta
      661 ttgcagcctg atatccttaa acgtctgcaa gaaatccaat ttgagcggat gaaaaagcag
      721 cgcaatgaga cgcagctgac agaaaaggaa gtcattgttc taaaagcaat tgctaaaggt
      781 cttaaaagca aagcgattgc ctttgatttg ggcgtctctg agcgaacagt aaagtccaga
      841 ttaacgtcca tttacaataa attaggcgcg aattcaagaa ctgaggcagt aacgattgcc
      901 atgcaaaaag gtattctgac aatagacaac taataataat ctagaccagg catcaaataa
      961 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1021 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatagtaaat
     1081 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg
     1141 acatcataac ggttctggca aatattctga aatgagctgg ttagctagtc aactgcatat
     1201 ttgaaaattg cccaaacgta catgcccgaa tgtacgtttt tttcatttca ttgtcaacta
     1261 caatgagaaa gaatgtgatc aagcaatgtg ttgaaaggag attatcaaag aggagaaata
     1321 ctagatgcgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg tggaactgga
     1381 tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa
     1441 tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac
     1501 tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg accatatgaa
     1561 gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc
     1621 ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct
     1681 ggtaaaccgc attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca
     1741 taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa
     1801 tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc
     1861 tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca
     1921 ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcatatggt
     1981 tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaatg
     2041 atgataataa tctagaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     2101 tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct
     2161 tcgggtgggc ctttctgcgt ttataggtac ccgtggatcc tctagagtcg acctgcaggc
     2221 atgcaagctt tttacggcta gctcagtcct aggtatagtg ctagcccagc cagagaaaca
     2281 ctctttaaca gggggtagta tgatgtctcg tttagataaa agtaaagtga ttaacagcgc
     2341 attagagctg cttaatgagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa
     2401 gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga
     2461 cgccttagcc attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga
     2521 aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca
     2581 tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct
     2641 cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc
     2701 actcagcgca gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt
     2761 cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc
     2821 tatcgaatta tttgatcacc aaggtgcaga gccagccttc ttattcggcc ttgaattgat
     2881 catatgcgga ttagaaaaac aacttaaatg tgaaagtggg tcttaaggca tcaaataaaa
     2941 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     3001 ctcctgagta ggacaaatcc gccgccctag acctagggcg ttcggctgcg gcgagcggta
     3061 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcatgcaag
     3121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3601 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     3661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3841 gtcatgacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct
     3901 gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga
     3961 gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     4021 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     4081 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     4141 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     4201 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccataacacg
     4261 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     4321 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     4381 atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc
     4441 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     4501 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     4561 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     4621 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaa
//
