LOCUS       sequence-247775-        4675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of narL(1-134aa)-ydfI(129-213aa) 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: pSR357.12
FEATURES             Location/Qualifiers
     source          1..4675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..116
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(137..155,156..161,162..178,179..184,185..210)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    137..155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    162..180
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      925..996
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1012..1039
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1278..1289
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1296..2009
                     /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
                     NFKIRHNVEDGSVQLADHYKQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2036..2107
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2123..2150
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2202..2236
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2271..2897
                     /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
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNTKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2347..2366)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2903..2989
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3154..3742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3234..3253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3830..3924)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3945..4604)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4519..4538
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggtgaaagtt ggaacctctt acgggccggc caacgtctca ttttcgccag atatcgacgt
       61 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
      121 tcgtcttcac ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga
      181 tactgagcac atcagcagga cgcactgacc taaggatttt ttctagaaga attatataca
      241 acaaatgagt aatcaggaac cggctactat cctgctgatt gacgatcacc cgatgctgcg
      301 aactggcgta aaacagctta tcagtatggc accagatatc accgtggttg gcgaagcgag
      361 taatggcgaa cagggtattg aactggcgga gtctcttgat cccgatctga tcctgttaga
      421 tctcaatatg cccggcatga acggtctgga aacgctggat aaactgcgcg aaaagtccct
      481 ctcagggcgc attgtggtat tcagcgtctc taaccatgaa gaagatgtgg tcaccgcact
      541 gaaacgcggc gcggatggct atctgttaaa agatatggaa ccggaagatc tgctgaaagc
      601 attgcatcag gcagctgctg gcgaaatggt attaagcgaa gcattactta aacgtctgca
      661 agaaatccaa tttgagcgga tgaaaaagca gcgcaatgag acgcagctga cagaaaagga
      721 agtcattgtt ctaaaagcaa ttgctaaagg tcttaaaagc aaagcgattg cctttgattt
      781 gggcgtctct gagcgaacag taaagtccag attaacgtcc atttacaata aattaggcgc
      841 gaattcaaga actgaggcag taacgattgc catgcaaaaa ggtattctga caatagacaa
      901 ctaataataa tctagaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
      961 tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct
     1021 tagggtgggc ctttctgcgt ttatagtaaa tcactgcata attcgtgtcg ctcaaggcgc
     1081 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
     1141 aaatgagctg gttagctagt caactgcata tttgaaaatt gcccaaacgt acatgcccga
     1201 atgtacgttt ttttcatttc attgtcaact acaatgagaa agaatgtgat caagcaatgt
     1261 gttgaaagga gattatcaaa gaggagaaat actagatgcg taaaggcgaa gagctgttca
     1321 ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg
     1381 tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta
     1441 ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc
     1501 agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc
     1561 cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc
     1621 gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg
     1681 actttaaaga agacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca
     1741 atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc
     1801 acaacgtgga ggatggcagc gtgcagctgg ctgatcacta caagcaaaac actccaatcg
     1861 gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta
     1921 aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca
     1981 tcacgcatgg tatggatgaa ctgtacaaat gatgataata atctagacca ggcatcaaat
     2041 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     2101 cgctatctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggta
     2161 ccagtggatc ctctagagtc gacctgcagg catgcaagct ttttacggct agctcagtcc
     2221 taggtatagt gctagcccag ccagagaaac actctttaac agggggtagt atgatgtctc
     2281 gtttagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg
     2341 aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt
     2401 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc
     2461 accatactca cttttgccct ttagaagggg aaagctggca agatttttta cgtaataaca
     2521 ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta
     2581 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac
     2641 aaggtttttc actagagaat gcattatatg cactcagcgc agtggggcat tttactttag
     2701 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta
     2761 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag
     2821 agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat
     2881 gtgaaagtgg gtcttaaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     2941 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta
     3001 gacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3061 atccacagaa tcaggggata acgcatgcaa gaacatgtga gcaaaaggcc agcaaaaggc
     3121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3781 gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa
     3841 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     3901 ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc
     3961 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     4021 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     4081 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     4141 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     4201 gggaaatagg ccaggttttc accataacac gccacatctt gcgaatatat gtgtagaaac
     4261 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     4321 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     4381 atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     4441 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     4501 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     4561 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     4621 cctgaaaatc tcgataactc aaaaaatacg accggtagtg atcttatttc attat
//
