LOCUS       sequence-247887-        4695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SO_4623(1-124aa)-psdR(134-237aa), ompR137 chimera 
            under PLtetO-1, output Promoter PpsdA110.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4695)
  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 4695)
  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: pKD285
FEATURES             Location/Qualifiers
     source          1..4695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..78
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(99..117,118..123,124..140,141..146,147..172)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    99..117
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    124..142
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      912..983
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      999..1026
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1260..1271
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1278..1991
                     /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      2018..2089
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2105..2132
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2184..2218
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2253..2879
                     /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(2329..2348)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2885..2971
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3136..3724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3216..3235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3812..3906)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3927..4586)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4501..4520
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cattttcgcc agatatcgac gtctaagaaa ccattattat catgacatta acctataaaa
       61 ataggcgtat cacgaggccc tttcgtcttc acctcgagtc cctatcagtg atagagattg
      121 acatccctat cagtgataga gatactgagc acatcagcag gacgcactga cccttaaatc
      181 ggcccctcat aagtcccttt tcacatgttg gtattactgg ttgaagacaa tcggcttctc
      241 tctaacaaca tcattcaata tcttgaattg agtggcatag agtgcgatta cgcgttcaac
      301 ctagcgcaag cagaaatgtt gatcagccag cagcaatttg acgccatcat ccttgatctt
      361 aatttaccgg atggtgatgg cattgaggcg tgtgaacgct ggaaagcgca atgttttagt
      421 tcacctatca tcatgctaac cgcccgtagc agtttgaatg agcgcttggc tggttttgct
      481 gttggcgctg atgattattt aataaaaccc tttgcgatgg aggagctggt cgcgcggctg
      541 aaggtggttg cccagcgtcg tcctgcgccc caaagggtgg agcttggcgg gctgacgatt
      601 tatcctgatc agaatgaggc cgaatggaac agcgtccgga ttttgttttc ccaaaaagaa
      661 ttccagcttc tctcgatttt tgtgagggag cataaaaaaa ttgtctctcg tgatgaacta
      721 ttggaggcgt tatgggatga cgttgatttt gtcgatgaca atacgttaac ggtcaatgtc
      781 aatcggctgc ggagaaaact ggagaatgca ggactgacag attgcatttc cacaatcaga
      841 ggacaaggtt atcaatttca agtcaatcgg aaggatgaag cggaatgtta ataataatct
      901 agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
      961 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     1021 tctgcgttta tagtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     1081 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctggtt
     1141 agctagtcac accctcgtga atgtgacagc attgtaagat tggggagcgg aattgcaaga
     1201 aagttcgatg ggaggatgct gacttccttt ttataataaa gaaaaaggag gagcagaaca
     1261 aagaggagaa atactagatg cgtaaaggcg aagagctgtt cactggtgtc gtccctattc
     1321 tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag
     1381 gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg
     1441 taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc
     1501 cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg
     1561 aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg
     1621 aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca
     1681 atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg
     1741 ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca
     1801 gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc
     1861 tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac
     1921 gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg
     1981 aactgtacaa atgatgataa taatctagac caggcatcaa ataaaacgaa aggctcagtc
     2041 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     2101 cactggctca ccttcgggtg ggcctttctg cgtttatagg tacccgtgga tcctctagag
     2161 tcgacctgca ggcatgcaag ctttttacgg ctagctcagt cctaggtata gtgctagccc
     2221 agccagagaa acactcttta acagggggta gtatgatgtc tcgtttagat aaaagtaaag
     2281 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta
     2341 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc
     2401 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc
     2461 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg
     2521 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac
     2581 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga
     2641 atgcattata tgcactcagc gcagtggggc attttacttt aggttgcgta ttggaagatc
     2701 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat
     2761 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg
     2821 gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtcttaag
     2881 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     2941 gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacctagg gcgttcggct
     3001 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3061 taacgcatgc aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3121 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3181 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3241 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3301 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3361 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3421 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3481 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3541 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3601 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3661 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3781 ttaagggatt ttggtcatga ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     3841 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     3901 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt
     3961 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     4021 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     4081 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     4141 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     4201 tcaccataac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     4261 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     4321 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     4381 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     4441 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     4501 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     4561 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     4621 tcaaaaaata cgaccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     4681 ggccggccaa cgtct
//
