LOCUS       sequence-247795-        4652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of uhpA(1-121aa)-ydfI(124-213aa), narL129 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: pSR322.3
FEATURES             Location/Qualifiers
     source          1..4652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..39
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(60..78,79..84,85..101,102..107,108..133)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    60..78
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    85..103
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      825..896
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      912..939
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1178..1189
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1196..1909
                     /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      1936..2007
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2023..2050
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2102..2136
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2171..2797
                     /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(2247..2266)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2803..2889
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3054..3642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3134..3153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3730..3824)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3845..4504)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4419..4438
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgagt
       61 ccctatcagt gatagagatt gacatcccta tcagtgatag agatactgag cacatcagca
      121 ggacgcactg accagttcta agacgtaaaa aagtagatag gacgccatat gatcaccgtt
      181 gcccttatag acgatcacct catcgtccgc tccggctttg cgcagctgct ggggctggaa
      241 cctgatttgc aggtagttgc cgagtttggt tcggggcgcg aggcgctggc ggggctgccg
      301 gggcgcggtg tgcaggtgtg tatttgcgat atctccatgc ccgatatctc cggtctggag
      361 ctgctaagcc agctgccgaa aggtatggcg acgattatgc tctccgttca cgacagtcct
      421 gcgctggttg agcaggcgct taacgcgggg gcacgcggct ttctttccaa acgctgtagc
      481 ccggatgaac tcattgctgc ggtgcatacg gttgccacgg gcggctgtta tttgcagcct
      541 gatatcctta aacgtctgca agaaatccaa tttgagcgga tgaaaaagca gcgcaatgag
      601 acgcagctga cagaaaagga agtcattgtt ctaaaagcaa ttgctaaagg tcttaaaagc
      661 aaagcgattg cctttgattt gggcgtctct gagcgaacag taaagtccag attaacgtcc
      721 atttacaata aattaggcgc gaattcaaga actgaggcag taacgattgc catgcaaaaa
      781 ggtattctga caatagacaa ctaataataa tctagaccag gcatcaaata aaacgaaagg
      841 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
      901 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatagtaaa tcactgcata
      961 attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa
     1021 cggttctggc aaatattctg aaatgagctg gttagctagt caactgcata tttgaaaatt
     1081 gcccaaacgt acatgcccga atgtacgttt ttttcatttc attgtcaact acaatgagaa
     1141 agaatgtgat caagcaatgt gttgaaagga gattatcaaa gaggagaaat actagatgcg
     1201 taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg atggtgatgt
     1261 caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta atggtaaact
     1321 gacgctgaag ttcatctgta ctactggtaa actgccggta ccttggccga ctctggtaac
     1381 gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga agcagcatga
     1441 cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt cctttaagga
     1501 tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg
     1561 cattgagctg aaaggcattg actttaaaga agacggcaat atcctgggcc ataagctgga
     1621 atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa atggcattaa
     1681 agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg ctgatcacta
     1741 ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc actatctgag
     1801 cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg ttctgctgga
     1861 gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat gatgataata
     1921 atctagacca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1981 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     2041 cctttctgcg tttataggta cccgtggatc ctctagagtc gacctgcagg catgcaagct
     2101 ttttacggct agctcagtcc taggtatagt gctagcccag ccagagaaac actctttaac
     2161 agggggtagt atgatgtctc gtttagataa aagtaaagtg attaacagcg cattagagct
     2221 gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt
     2281 agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc
     2341 cattgagatg ttagataggc accatactca cttttgccct ttagaagggg aaagctggca
     2401 agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg
     2461 agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca
     2521 attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc
     2581 agtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga
     2641 agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt
     2701 atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg
     2761 attagaaaaa caacttaaat gtgaaagtgg gtcttaaggc atcaaataaa acgaaaggct
     2821 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     2881 aggacaaatc cgccgcccta gacctagggc gttcggctgc ggcgagcggt atcagctcac
     2941 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcatgcaa gaacatgtga
     3001 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3061 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3121 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3181 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3241 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3301 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3361 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3421 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3481 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3541 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3601 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3661 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgact
     3721 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     3781 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat
     3841 caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     3901 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     3961 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     4021 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     4081 tattctcaat aaacccttta gggaaatagg ccaggttttc accataacac gccacatctt
     4141 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     4201 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     4261 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
     4321 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     4381 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     4441 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     4501 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg accggtagtg
     4561 atcttatttc attatggtga aagttggaac ctcttacggg ccggccaacg tctcattttc
     4621 gccagatatc gacgtctaag aaaccattat ta
//
