LOCUS       sequence-239147-        4624 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 4624)
  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 4624)
  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..4624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(596..690)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(711..1370)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1285..1304
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1371..1473)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1539..1557
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1578..1596,1597..1602,1603..1619,1620..1625,
                     1626..1651)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1578..1596
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1603..1621
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2316..2387
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2403..2430
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2669..2680
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2687..3400
                     /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      3427..3498
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3514..3541
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3593..3627
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             3662..4288
                     /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(3738..3757)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      4294..4380
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(join(4544..4624,1..508))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(join(4624,1..19))
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
       61 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
      121 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      181 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      241 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      301 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      361 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      421 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      481 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      541 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgactagtg cttggattct
      601 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
      661 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg
      721 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
      781 tcacagacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
      841 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
      901 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
      961 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     1021 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     1081 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     1141 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     1201 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     1261 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     1321 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     1381 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     1441 tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca gatatcgacg
     1501 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     1561 ttcgtcttca cctcgagtcc ctatcagtga tagagattga catccctatc agtgatagag
     1621 atactgagca catcagcagg acgcactgac cagttctaag acgtaaaaaa gtagatagga
     1681 cgccatatga tcaccgttgc ccttatagac gatcacctca tcgtccgctc cggctttgcg
     1741 cagctgctgg ggctggaacc tgatttgcag gtagttgccg agtttggttc ggggcgcgag
     1801 gcgctggcgg ggctgccggg gcgcggtgtg caggtgtgta tttgcgatat ctccatgccc
     1861 gatatctccg gtctggagct gctaagccag ctgccgaaag gtatggcgac gattatgctc
     1921 tccgttcacg acagtcctgc gctggttgag caggcgctta acgcgggggc acgcggcttt
     1981 ctttccaaac gctgtagccc ggatgaactc attgctgcgg tgcatacggt tgccacgggc
     2041 ggctgttatc tgacgccgga tattgccatt aaactggcat ccggtcgtca ggacccgcta
     2101 accaaacgtg aagtcattgt tctaaaagca attgctaaag gtcttaaaag caaagcgatt
     2161 gcctttgatt tgggcgtctc tgagcgaaca gtaaagtcca gattaacgtc catttacaat
     2221 aaattaggcg cgaattcaag aactgaggca gtaacgattg ccatgcaaaa aggtattctg
     2281 acaatagaca actaataata atctagacca ggcatcaaat aaaacgaaag gctcagtcga
     2341 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     2401 ctggctcacc ttcgggtggg cctttctgcg tttatagtaa atcactgcat aattcgtgtc
     2461 gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg
     2521 caaatattct gaaatgagct ggttagctag tcaactgcat atttgaaaat tgcccaaacg
     2581 tacatgcccg aatgtacgtt tttttcattt cattgtcaac tacaatgaga aagaatgtga
     2641 tcaagcaatg tgttgaaagg agattatcaa agaggagaaa tactagatgc gtaaaggcga
     2701 agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg tcaacggtca
     2761 taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac tgacgctgaa
     2821 gttcatctgt actactggta aactgccggt accttggccg actctggtaa cgacgctgac
     2881 ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg acttcttcaa
     2941 gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg atgacggcac
     3001 gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc gcattgagct
     3061 gaaaggcatt gactttaaag aagacggcaa tatcctgggc cataagctgg aatacaattt
     3121 taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta aagcgaattt
     3181 taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact accagcaaaa
     3241 cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga gcacgcaaag
     3301 cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg agttcgtaac
     3361 cgcagcgggc atcacgcatg gtatggatga actgtacaaa tgatgataat aatctagacc
     3421 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3481 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     3541 gtttataggt acccgtggat cctctagagt cgacctgcag gcatgcaagc tttttacggc
     3601 tagctcagtc ctaggtatag tgctagccca gccagagaaa cactctttaa cagggggtag
     3661 tatgatgtct cgtttagata aaagtaaagt gattaacagc gcattagagc tgcttaatga
     3721 ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg tagagcagcc
     3781 tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag ccattgagat
     3841 gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc aagatttttt
     3901 acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg gagcaaaagt
     3961 acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc aattagcctt
     4021 tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg cagtggggca
     4081 ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag aagaaaggga
     4141 aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat tatttgatca
     4201 ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg gattagaaaa
     4261 acaacttaaa tgtgaaagtg ggtcttaagg catcaaataa aacgaaaggc tcagtcgaaa
     4321 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     4381 ccgccgccct agacctaggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4441 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4501 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4561 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4621 tata
//
