LOCUS       sequence-108027-        3936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CcaR constitutively and sfGFP under the PcpcG2-172 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 35)
  AUTHORS   Schmidl SR, Sheth RU, Wu A, Tabor JJ
  TITLE     Refactoring and optimization of light-switchable escherichia coli 
            two-component systems.
  JOURNAL   ACS Synth Biol. 2014 Nov 21;3(11):820-31. doi: 10.1021/sb500273n. 
            Epub 2014 Oct 11.
  PUBMED    25250630
REFERENCE   2  (bases 1 to 3936)
  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: "10.1021/sb500273n";
            journalName: "ACS Synth Biol"; date: "2014-11-21- 21"; volume: "3"; 
            issue: "11"; pages: "820-31"
COMMENT     Sequence Label: pSR58.6
FEATURES             Location/Qualifiers
     source          1..3936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     terminator      795..866
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      882..909
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1205..1216
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1223..1936
                     /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      1963..2034
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2050..2077
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2279..2867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2359..2378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2955..3049)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3070..3729)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3644..3663
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3730..3832)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3898..3916
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttgacggcta gctcagtcct aggtacagtg ctagctgtta aacgctcctc gtaagaacac
       61 gggaaagcaa tgagaattct tttagtggag gatgatttgc cgctggcgga aacccttgct
      121 gaagcattga gtgaccagct ttacaccgtt gatattgcca ccgacgcttc cctcgcctgg
      181 gactatgcct cccgactgga atatgacctc gttattttgg atgtgatgct gccggagttg
      241 gacgggatta ccctctgtca aaaatggcga tcgcacagtt atttaatgcc aattttgatg
      301 atgacagcca gggatacgat caatgataaa atcacgggct tggatgcggg ggcggatgat
      361 tatgtggtca agccagtgga tttgggggag ttatttgcca gggtgcgagc tttgttgcgt
      421 cggggttgtg caacgtgcca accagtttta gagtgggggc caatcaggtt ggatccaagc
      481 acctatgaag ttagttatga caatgaggtt ttgtctttga cccgcaagga atacagcatt
      541 ctggaattac tactccgcaa tggccgtcgg gtgctaagtc ggagcatgat tatcgatagt
      601 atctggaagt tggagagtcc cccagaggaa gatacggtta aggtgcatgt gcggagtttg
      661 cgacaaaaat taaaaagtgc cggtttatca gcagatgcca ttgaaacggt ccatggcatt
      721 gggtatcgtc tggccaattt aacggaaaaa tctttgtgcc aagggaaaaa ctagtaataa
      781 tctagaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      841 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
      901 ctttctgcgt ttatagtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc
      961 tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
     1021 gttagctagt caagcccatt gtgcttttct ctatcaacct cagcttacct gaaggggtga
     1081 acaggtctgg gttaattcat gttgcgaaat gtaacagttt tagtcgcatc agctaacttt
     1141 ccgatttctt tacgattttc tccccctttt cttcaatttt actttgttag gatcgcattt
     1201 ttaaaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc
     1261 tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg
     1321 tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact
     1381 gccggtacct tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg
     1441 ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt
     1501 gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa
     1561 atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagaaga
     1621 cggcaatatc ctgggccata agctggaata caattttaac agccacaatg tttacatcac
     1681 cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga
     1741 tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt
     1801 tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga
     1861 gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat
     1921 ggatgaactg tacaaatgat gataataatc tagaccaggc atcaaataaa acgaaaggct
     1981 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     2041 agtcacactg gctcaccttc gggtgggcct ttctgcgttt ataggtaccc gtggatcctc
     2101 tagagtcgac ctgcaggcat gcaagcttcc taggcgttcg gctgcggcga gcggtatcag
     2161 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca tgcaagaaca
     2221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2461 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2701 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2941 tgactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
     3001 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
     3061 gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     3121 tctgccgaca tggaagccat cacagacggc atgatgaacc tgaatcgcca gcggcatcag
     3181 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     3241 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     3301 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     3361 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     3421 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     3481 caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc
     3541 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     3601 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     3661 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     3721 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     3781 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     3841 ttttcgccag atatcgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     3901 aggcgtatca cgaggccctt tcgtcttcac ctcgag
//
