LOCUS       sequence-247799-        4625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of uhpA(1-141aa)-ydfI(153-213aa), narL159 chimera under 
            PLtetO-1, output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4625)
  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 4625)
  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: pSR322.4
FEATURES             Location/Qualifiers
     source          1..4625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..46
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(67..85,86..91,92..108,109..114,115..140)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    67..85
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    92..110
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      805..876
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      892..919
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1158..1169
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1176..1889
                     /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      1916..1987
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2003..2030
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2082..2116
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2151..2777
                     /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(2227..2246)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2783..2869
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3034..3622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3114..3133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3710..3804)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3825..4484)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4399..4418
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
       61 ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
      121 atcagcagga cgcactgacc agttctaaga cgtaaaaaag tagataggac gccatatgat
      181 caccgttgcc cttatagacg atcacctcat cgtccgctcc ggctttgcgc agctgctggg
      241 gctggaacct gatttgcagg tagttgccga gtttggttcg gggcgcgagg cgctggcggg
      301 gctgccgggg cgcggtgtgc aggtgtgtat ttgcgatatc tccatgcccg atatctccgg
      361 tctggagctg ctaagccagc tgccgaaagg tatggcgacg attatgctct ccgttcacga
      421 cagtcctgcg ctggttgagc aggcgcttaa cgcgggggca cgcggctttc tttccaaacg
      481 ctgtagcccg gatgaactca ttgctgcggt gcatacggtt gccacgggcg gctgttatct
      541 gacgccggat attgccatta aactggcatc cggtcgtcag gacccgctaa ccaaacgtga
      601 agtcattgtt ctaaaagcaa ttgctaaagg tcttaaaagc aaagcgattg cctttgattt
      661 gggcgtctct gagcgaacag taaagtccag attaacgtcc atttacaata aattaggcgc
      721 gaattcaaga actgaggcag taacgattgc catgcaaaaa ggtattctga caatagacaa
      781 ctaataataa tctagaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
      841 tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct
      901 tcgggtgggc ctttctgcgt ttatagtaaa tcactgcata attcgtgtcg ctcaaggcgc
      961 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
     1021 aaatgagctg gttagctagt caactgcata tttgaaaatt gcccaaacgt acatgcccga
     1081 atgtacgttt ttttcatttc attgtcaact acaatgagaa agaatgtgat caagcaatgt
     1141 gttgaaagga gattatcaaa gaggagaaat actagatgcg taaaggcgaa gagctgttca
     1201 ctggtgtcgt ccctattctg gtggaactgg atggtgatgt caacggtcat aagttttccg
     1261 tgcgtggcga gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta
     1321 ctactggtaa actgccggta ccttggccga ctctggtaac gacgctgact tatggtgttc
     1381 agtgctttgc tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc
     1441 cggaaggcta tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc
     1501 gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg
     1561 actttaaaga agacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca
     1621 atgtttacat caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc
     1681 acaacgtgga ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg
     1741 gtgatggtcc tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta
     1801 aagatccgaa cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca
     1861 tcacgcatgg tatggatgaa ctgtacaaat gatgataata atctagacca ggcatcaaat
     1921 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1981 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggta
     2041 cccgtggatc ctctagagtc gacctgcagg catgcaagct ttttacggct agctcagtcc
     2101 taggtatagt gctagcccag ccagagaaac actctttaac agggggtagt atgatgtctc
     2161 gtttagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg
     2221 aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt
     2281 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc
     2341 accatactca cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg
     2401 ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta
     2461 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac
     2521 aaggtttttc actagagaat gcattatatg cactcagcgc agtggggcat tttactttag
     2581 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta
     2641 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag
     2701 agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat
     2761 gtgaaagtgg gtcttaaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     2821 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta
     2881 gacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2941 atccacagaa tcaggggata acgcatgcaa gaacatgtga gcaaaaggcc agcaaaaggc
     3001 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3061 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3121 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3181 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3241 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3301 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3361 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3421 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3481 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3541 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3601 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3661 gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa
     3721 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     3781 ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc
     3841 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     3901 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     3961 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     4021 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     4081 gggaaatagg ccaggttttc accataacac gccacatctt gcgaatatat gtgtagaaac
     4141 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     4201 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     4261 atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     4321 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     4381 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     4441 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     4501 cctgaaaatc tcgataactc aaaaaatacg accggtagtg atcttatttc attatggtga
     4561 aagttggaac ctcttacggg ccggccaacg tctcattttc gccagatatc gacgtctaag
     4621 aaacc
//
