LOCUS       sequence-247890-        4707 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of SO_4428(1-127aa)-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 4707)
  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 4707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pKD281
FEATURES             Location/Qualifiers
     source          1..4707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(27..45,46..51,52..68,69..74,75..100)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    27..45
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    52..70
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      852..923
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      939..966
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1200..1211
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1218..1931
                     /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      1958..2029
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2045..2072
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2124..2158
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2193..2819
                     /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(2269..2288)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2825..2911
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3076..3664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3156..3175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3752..3846)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3867..4526)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4441..4460
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     join(4695..4707,1..6)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgaggccctt tcgtcttcac ctcgagtccc tatcagtgat agagattgac atccctatca
       61 gtgatagaga tactgagcac atcagcagga cgcactgacc tacccagccc agtaagtaca
      121 tattagaaag ggttaatgaa aatactttta gttgaagatg atgcgacaac gattgattac
      181 atcgttaaag gatttcttga gcagggacat aatattgaaa ccgccagtga tggtcaccaa
      241 ggactactgc aagccacgag tggccagtat gatctgttga tcctcgaccg catgttgcct
      301 caactcgatg gtttaaaact cttggccgcc ctcagggcta ctggcaatca aaccccagta
      361 ttgatcctgt cggcgcttgc ccatgtggat gaacgggtaa aaggtctgcg cgcaggcggc
      421 gatgattata tgacaaagcc ctttgctttt tcggagctgt tagtgcgtgc cgaaaagctg
      481 atgcaacggg gccaatccac gcccattacc acagatgtgg agcttggcgg gctgacgatt
      541 tatcctgatc agaatgaggc cgaatggaac agcgtccgga ttttgttttc ccaaaaagaa
      601 ttccagcttc tctcgatttt tgtgagggag cataaaaaaa ttgtctctcg tgatgaacta
      661 ttggaggcgt tatgggatga cgttgatttt gtcgatgaca atacgttaac ggtcaatgtc
      721 aatcggctgc ggagaaaact ggagaatgca ggactgacag attgcatttc cacaatcaga
      781 ggacaaggtt atcaatttca agtcaatcgg aaggatgaag cggaatgtta ataataatct
      841 agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
      901 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
      961 tctgcgttta tagtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     1021 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctggtt
     1081 agctagtcac accctcgtga atgtgacagc attgtaagat tggggagcgg aattgcaaga
     1141 aagttcgatg ggaggatgct gacttccttt ttataataaa gaaaaaggag gagcagaaca
     1201 aagaggagaa atactagatg cgtaaaggcg aagagctgtt cactggtgtc gtccctattc
     1261 tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag
     1321 gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg
     1381 taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc
     1441 cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg
     1501 aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg
     1561 aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca
     1621 atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg
     1681 ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca
     1741 gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc
     1801 tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac
     1861 gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg
     1921 aactgtacaa atgatgataa taatctagac caggcatcaa ataaaacgaa aggctcagtc
     1981 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     2041 cactggctca ccttcgggtg ggcctttctg cgtttatagg tacccgtgga tcctctagag
     2101 tcgacctgca ggcatgcaag ctttttacgg ctagctcagt cctaggtata gtgctagccc
     2161 agccagagaa acactcttta acagggggta gtatgatgtc tcgtttagat aaaagtaaag
     2221 tgattaacag cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta
     2281 aactcgccca gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc
     2341 gggctttgct cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc
     2401 ctttagaagg ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg
     2461 ctttactaag tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac
     2521 agtatgaaac tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga
     2581 atgcattata tgcactcagc gcagtggggc attttacttt aggttgcgta ttggaagatc
     2641 aagagcatca agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat
     2701 tattacgaca agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg
     2761 gccttgaatt gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtcttaag
     2821 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     2881 gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacctagg gcgttcggct
     2941 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3001 taacgcatgc aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3061 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3121 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3181 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3241 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3301 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3361 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3421 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3481 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3541 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3601 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3661 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3721 ttaagggatt ttggtcatga ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     3781 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     3841 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt
     3901 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     3961 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     4021 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     4081 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     4141 tcaccataac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     4201 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     4261 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     4321 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     4381 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     4441 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     4501 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     4561 tcaaaaaata cgaccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     4621 ggccggccaa cgtctcattt tcgccagata tcgacgtcta agaaaccatt attatcatga
     4681 cattaaccta taaaaatagg cgtatca
//
