LOCUS       sequence-251168-        4651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of uhpA(1-118aa)-ydfI(121-213aa), narL126 chimera under 
            PLtetO-1, output Promoter PydfJ115.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4651)
  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 4651)
  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: pSR322.2
FEATURES             Location/Qualifiers
     source          1..4651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..42
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(63..81,82..87,88..104,105..110,111..136)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    63..81
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    88..106
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      828..899
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      915..942
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1180..1191
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1198..1911
                     /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      1938..2009
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2025..2052
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2104..2138
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2173..2799
                     /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(2249..2268)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2805..2891
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3056..3644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3136..3155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3732..3826)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3847..4506)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4421..4440
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg
       61 agtccctatc agtgatagag attgacatcc ctatcagtga tagagatact gagcacatca
      121 gcaggacgca ctgaccagtt ctaagacgta aaaaagtaga taggacgcca tatgatcacc
      181 gttgccctta tagacgatca cctcatcgtc cgctccggct ttgcgcagct gctggggctg
      241 gaacctgatt tgcaggtagt tgccgagttt ggttcggggc gcgaggcgct ggcggggctg
      301 ccggggcgcg gtgtgcaggt gtgtatttgc gatatctcca tgcccgatat ctccggtctg
      361 gagctgctaa gccagctgcc gaaaggtatg gcgacgatta tgctctccgt tcacgacagt
      421 cctgcgctgg ttgagcaggc gcttaacgcg ggggcacgcg gctttctttc caaacgctgt
      481 agcccggatg aactcattgc tgcggtgcat acggttgcca cgggcaatgt gctattgcag
      541 cctgatatcc ttaaacgtct gcaagaaatc caatttgagc ggatgaaaaa gcagcgcaat
      601 gagacgcagc tgacagaaaa ggaagtcatt gttctaaaag caattgctaa aggtcttaaa
      661 agcaaagcga ttgcctttga tttgggcgtc tctgagcgaa cagtaaagtc cagattaacg
      721 tccatttaca ataaattagg cgcgaattca agaactgagg cagtaacgat tgccatgcaa
      781 aaaggtattc tgacaataga caactaataa taatctagac caggcatcaa ataaaacgaa
      841 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
      901 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagt aaatcactgc
      961 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca
     1021 taacggttct ggcaaatatt ctgaaatgag ctggttagct agtcaactgc atatttaaaa
     1081 ttgcccaaac gtacatgccc gaatgtacgt ttttttcatt tcattgtcaa ctacaatgag
     1141 aaagaatgtg atcaagcaat gtgttgaaag gagattatca aagaggagaa atactagatg
     1201 cgtaaaggcg aagagctgtt cactggtgtc gtccctattc tggtggaact ggatggtgat
     1261 gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa
     1321 ctgacgctga agttcatctg tactactggt aaactgccgg taccttggcc gactctggta
     1381 acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat
     1441 gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag
     1501 gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac
     1561 cgcattgagc tgaaaggcat tgactttaaa gaagacggca atatcctggg ccataagctg
     1621 gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt
     1681 aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac
     1741 taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg
     1801 agcacgcaaa gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg
     1861 gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa atgatgataa
     1921 taatctagac caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1981 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     2041 ggcctttctg cgtttatagg tacccgtgga tcctctagag tcgacctgca ggcatgcaag
     2101 ctttttacgg ctagctcagt cctaggtata gtgctagccc agccagagaa acactcttta
     2161 acagggggta gtatgatgtc tcgtttagat aaaagtaaag tgattaacag cgcattagag
     2221 ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt
     2281 gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta
     2341 gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg
     2401 caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat
     2461 ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat
     2521 caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc
     2581 gcagtggggc attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa
     2641 gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa
     2701 ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatatgc
     2761 ggattagaaa aacaacttaa atgtgaaagt gggtcttaag gcatcaaata aaacgaaagg
     2821 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     2881 gtaggacaaa tccgccgccc tagacctagg gcgttcggct gcggcgagcg gtatcagctc
     2941 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcatgc aagaacatgt
     3001 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3481 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3721 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     3781 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat
     3841 atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     3901 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     3961 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     4021 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     4081 catattctca ataaaccctt tagggaaata ggccaggttt tcaccataac acgccacatc
     4141 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     4201 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     4261 cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag
     4321 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     4381 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     4441 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     4501 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgaccggtag
     4561 tgatcttatt tcattatggt gaaagttgga acctcttacg ggccggccaa cgtctcattt
     4621 tcgccagata tcgacgtcta agaaaccatt a
//
