LOCUS       Exported                3936 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses CcaR constitutively and sfGFP under the PcpcG2-172 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pSR58.6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3936)
  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
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"
FEATURES             Location/Qualifiers
     source          1..3936
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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      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"