LOCUS       sequence-245253-        4652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of uhpA(1-123aa)-ydfI(126-213aa), narL131 chimera under 
            PLtetO-1, output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4652)
  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 4652)
  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: pSR322
FEATURES             Location/Qualifiers
     source          1..4652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     51..69
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(90..108,109..114,115..131,132..137,138..163)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    90..108
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    115..133
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      855..926
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      942..969
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1208..1219
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1226..1939
                     /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      1966..2037
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2053..2080
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2132..2166
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2201..2827
                     /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(2277..2296)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2833..2919
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3084..3672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3164..3183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3760..3854)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3875..4534)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4449..4468
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cagatatcga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
       61 tcacgaggcc ctttcgtctt cacctcgagt ccctatcagt gatagagatt gacatcccta
      121 tcagtgatag agatactgag cacatcagca ggacgcactg accagttcta agacgtaaaa
      181 aagtagatag gacgccatat gatcaccgtt gcccttatag acgatcacct catcgtccgc
      241 tccggctttg cgcagctgct ggggctggaa cctgatttgc aggtagttgc cgagtttggt
      301 tcggggcgcg aggcgctggc ggggctgccg gggcgcggtg tgcaggtgtg tatttgcgat
      361 atctccatgc ccgatatctc cggtctggag ctgctaagcc agctgccgaa aggtatggcg
      421 acgattatgc tctccgttca cgacagtcct gcgctggttg agcaggcgct taacgcgggg
      481 gcacgcggct ttctttccaa acgctgtagc ccggatgaac tcattgctgc ggtgcatacg
      541 gttgccacgg gcggctgtta tctgacgcct gatatcctta aacgtctgca agaaatccaa
      601 tttgagcgga tgaaaaagca gcgcaatgag acgcagctga cagaaaagga agtcattgtt
      661 ctaaaagcaa ttgctaaagg tcttaaaagc aaagcgattg cctttgattt gggcgtctct
      721 gagcgaacag taaagtccag attaacgtcc atttacaata aattaggcgc gaattcaaga
      781 actgaggcag taacgattgc catgcaaaaa ggtattctga caatagacaa ctaataataa
      841 tctagaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      901 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
      961 ctttctgcgt ttatagtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc
     1021 tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
     1081 gttagctagt caactgcata tttgaaaatt gcccaaacgt acatgcccga atgtacgttt
     1141 ttttcatttc attgtcaact acaatgagaa agaatgtgat caagcaatgt gttgaaagga
     1201 gattatcaaa gaggagaaat actagatgcg taaaggcgaa gagctgttca ctggtgtcgt
     1261 ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg tgcgtggcga
     1321 gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa
     1381 actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc
     1441 tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta
     1501 tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt
     1561 gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga
     1621 agacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca atgtttacat
     1681 caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc acaacgtgga
     1741 ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg gtgatggtcc
     1801 tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta aagatccgaa
     1861 cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg
     1921 tatggatgaa ctgtacaaat gatgataata atctagacca ggcatcaaat aaaacgaaag
     1981 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     2041 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggta cccgtggatc
     2101 ctctagagtc gacctgcagg catgcaagct ttttacggct agctcagtcc taggtatagt
     2161 gctagcccag ccagagaaac actctttaac agggggtagt atgatgtctc gtttagataa
     2221 aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac
     2281 aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa
     2341 aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc accatactca
     2401 cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt
     2461 tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac
     2521 agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc
     2581 actagagaat gcattatatg cactcagcgc agtggggcat tttactttag gttgcgtatt
     2641 ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat
     2701 gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt
     2761 cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg
     2821 gtcttaaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2881 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagggc
     2941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3001 tcaggggata acgcatgcaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3481 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     3541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3721 aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa taaaaaacgc
     3781 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     3841 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
     3901 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     3961 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     4021 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     4081 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     4141 ccaggttttc accataacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     4201 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     4261 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
     4321 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     4381 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     4441 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     4501 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     4561 tcgataactc aaaaaatacg accggtagtg atcttatttc attatggtga aagttggaac
     4621 ctcttacggg ccggccaacg tctcattttc gc
//
