LOCUS       sequence-245240-        4728 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of SO_2193(1-134aa)-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 4728)
  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 4728)
  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: pKD277
FEATURES             Location/Qualifiers
     source          1..4728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..50
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(71..89,90..95,96..112,113..118,119..144)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    71..89
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    96..114
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      917..988
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1004..1031
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1265..1276
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1283..1996
                     /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      2023..2094
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2110..2137
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2189..2223
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             2258..2884
                     /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(2334..2353)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2890..2976
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3141..3729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3221..3240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3817..3911)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3932..4591)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4506..4525
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
       61 tcacctcgag tccctatcag tgatagagat tgacatccct atcagtgata gagatactga
      121 gcacatcagc aggacgcact gacccgaaag agtaaacagc ccgacacggt aagtagccta
      181 tgaaacgtat cgctattgtg gaagatgaag ccgccattcg cgaaaattac aaggatgtgc
      241 tgcagcaaca cggttacagt gtccaaacct atgccgaccg gccatcggcc atgttggcgt
      301 ttaatacgcg cttaccggat ttagccatta tcgatatcgg ccttggcaat gagatcgacg
      361 gtggatttat gctgtgtcag tccctacgtg ccatgtcaaa caccttaccg attattttct
      421 taaccgctcg ggatagcgat ttcgatactg tatgtggcct tcgtttaggt gccgatgatt
      481 acctgagcaa agaagtcagt tttccccatt tgacggcgcg tttagccgcc ctttttcgac
      541 gctcagaact cgccgccagc caaacaccgc aggaaaatct agtggagctt ggcgggctga
      601 cgatttatcc tgatcagaat gaggccgaat ggaacagcgt ccggattttg ttttcccaaa
      661 aagaattcca gcttctctcg atttttgtga gggagcataa aaaaattgtc tctcgtgatg
      721 aactattgga ggcgttatgg gatgacgttg attttgtcga tgacaatacg ttaacggtca
      781 atgtcaatcg gctgcggaga aaactggaga atgcaggact gacagattgc atttccacaa
      841 tcagaggaca aggttatcaa tttcaagtca atcggaagga tgaagcggaa tgttaataat
      901 aatctagacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
      961 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     1021 gcctttctgc gtttatagta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     1081 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     1141 tggttagcta gtcacaccct cgtgaatgtg acagcattgt aagattgggg agcggaattg
     1201 caagaaagtt cgatgggagg atgctgactt cctttttata ataaagaaaa aggaggagca
     1261 gaacaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc
     1321 tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg
     1381 tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact
     1441 gccggtacct tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg
     1501 ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt
     1561 gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa
     1621 atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagaaga
     1681 cggcaatatc ctgggccata agctggaata caattttaac agccacaatg tttacatcac
     1741 cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga
     1801 tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt
     1861 tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga
     1921 gaaacgcgat catatggttc tgctggagtt cgtgaccgca gcgggcatca cgcatggtat
     1981 ggatgaactg tacaaatgat gataataatc tagaccaggc atcaaataaa acgaaaggct
     2041 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     2101 agtcacactg gctcaccttc gggtgggcct ttctgcgttt ataggtaccc gtggatcctc
     2161 tagagtcgac ctgcaggcat gcaagctttt tacggctagc tcagtcctag gtatagtgct
     2221 agcccagcca gagaaacact ctttaacagg gggtagtatg atgtctcgtt tagataaaag
     2281 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac
     2341 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa
     2401 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt
     2461 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag
     2521 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga
     2581 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact
     2641 agagaatgca ttatatgcac tcagcgcagt ggggcatttt actttaggtt gcgtattgga
     2701 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc
     2761 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt
     2821 attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc
     2881 ttaaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2941 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagggcgtt
     3001 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3061 ggggataacg catgcaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3121 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3181 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3241 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3301 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3361 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3421 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3481 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3541 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3601 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3661 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3721 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3781 tcacgttaag ggattttggt catgactagt gcttggattc tcaccaataa aaaacgcccg
     3841 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     3901 caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt
     3961 actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa
     4021 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
     4081 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     4141 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     4201 ggttttcacc ataacacgcc acatcttgcg aatatatgtg tagaaactgt cggaaatcgt
     4261 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     4321 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
     4381 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     4441 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     4501 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     4561 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     4621 ataactcaaa aaatacgacc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     4681 ttacgggccg gccaacgtct cattttcgcc agatatcgac gtctaaga
//
