LOCUS       sequence-247883-        4674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-120aa)-ydfI(115-213aa) chimera under PLtetO-1, 
            output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4674)
  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 4674)
  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: pSR357.27
FEATURES             Location/Qualifiers
     source          1..4674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(34..52,53..58,59..75,76..81,82..107)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    34..52
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    59..77
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      822..893
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      909..936
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1175..1186
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1193..1906
                     /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      1933..2004
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2020..2047
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2099..2133
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2168..2794
                     /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(2244..2263)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2800..2886
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3050..3638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3130..3149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3726..3820)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3841..4500)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4415..4434
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     join(4669..4674,1..13)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtatcacga ggccctttcg tcttcacctc gagtccctat cagtgataga gattgacatc
       61 cctatcagtg atagagatac tgagcacatc agcaggacgc actgacctaa ggattttttc
      121 tagaagaatt atatacaaca aatgagtaat caggaaccgg ctactatcct gctgattgac
      181 gatcacccga tgctgcgaac tggcgtaaaa cagcttatca gtatggcacc agatatcacc
      241 gtggttggcg aagcgagtaa tggcgaacag ggtattgaac tggcggagtc tcttgatccc
      301 gatctgatcc tgttagatct caatatgccc ggcatgaacg gtctggaaac gctggataaa
      361 ctgcgcgaaa agtccctctc agggcgcatt gtggtattca gcgtctctaa ccatgaagaa
      421 gatgtggtca ccgcactgaa acgcggcgcg gatggctatc tgttaaaaga tatggaaccg
      481 gaagatctgc tgaaagcatt ggatgcagcc atcagaggaa atgtgctatt gcagcctgat
      541 atccttaaac gtctgcaaga aatccaattt gagcggatga aaaagcagcg caatgagacg
      601 cagctgacag aaaaggaagt cattgttcta aaagcaattg ctaaaggtct taaaagcaaa
      661 gcgattgcct ttgatttggg cgtctctgag cgaacagtaa agtccagatt aacgtccatt
      721 tacaataaat taggcgcgaa ttcaagaact gaggcagtaa cgattgccat gcaaaaaggt
      781 attctgacaa tagacaacta ataataatct agaccaggca tcaaataaaa cgaaaggctc
      841 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
      901 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tagtaaatca ctgcataatt
      961 cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg
     1021 ttctggcaaa tattctgaaa tgagctggtt agctagtcaa ctgcatattt gaaaattgcc
     1081 caaacgtaca tgcccgaatg tacgtttttt tcatttcatt gtcaactaca atgagaaaga
     1141 atgtgatcaa gcaatgtgtt gaaaggagat tatcaaagag gagaaatact agatgcgtaa
     1201 aggcgaagag ctgttcactg gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa
     1261 cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac
     1321 gctgaagttc atctgtacta ctggtaaact gccggtacct tggccgactc tggtaacgac
     1381 gctgacttat ggtgttcagt gctttgctcg ttatccggac catatgaagc agcatgactt
     1441 cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga
     1501 cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg taaaccgcat
     1561 tgagctgaaa ggcattgact ttaaagaaga cggcaatatc ctgggccata agctggaata
     1621 caattttaac agccacaatg tttacatcac cgccgataaa caaaaaaatg gcattaaagc
     1681 gaattttaaa attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca
     1741 gcaaaacact ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac
     1801 gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat catatggttc tgctggagtt
     1861 cgtaaccgca gcgggcatca cgcatggtat ggatgaactg tacaaatgat gataataatc
     1921 tagaccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1981 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     2041 ttctgcgttt ataggtaccc gtggatcctc tagagtcgac ctgcaggcat gcaagctttt
     2101 tacggctagc tcagtcctag gtatagtgct agcccagcca gagaaacact ctttaacagg
     2161 gggtagtatg atgtctcgtt tagataaaag taaagtgatt aacagcgcat tagagctgct
     2221 taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga
     2281 gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat
     2341 tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga
     2401 ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc
     2461 aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt
     2521 agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgcagt
     2581 ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga
     2641 aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt
     2701 tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt
     2761 agaaaaacaa cttaaatgtg aaagtgggtc ttaaggcatc aaataaaacg aaaggctcag
     2821 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2881 acaaatccgc cgccctagac ctagggcgtt cggctgcggc gagcggtatc agctcatcaa
     2941 aggcggtaat acggttatcc acagaatcag gggataacgc atgcaagaac atgtgagcaa
     3001 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3061 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3121 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3181 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3241 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3301 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3361 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3421 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3481 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3661 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgactagtg
     3721 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     3781 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     3841 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     3901 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     3961 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     4021 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     4081 ctcaataaac cctttaggga aataggccag gttttcacca taacacgcca catcttgcga
     4141 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     4201 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     4261 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
     4321 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     4381 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     4441 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     4501 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgaccg gtagtgatct
     4561 tatttcatta tggtgaaagt tggaacctct tacgggccgg ccaacgtctc attttcgcca
     4621 gatatcgacg tctaagaaac cattattatc atgacattaa cctataaaaa tagg
//
