LOCUS       sequence-79911-e        6443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CcaS and CcaR constitutively, and sfGFP under the PcpcG2 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  AUTHORS   Olson EJ, Hartsough LA, Landry BP, Shroff R, Tabor JJ
  TITLE     Characterizing bacterial gene circuit dynamics with optically 
            programmed gene expression signals.
  JOURNAL   Nat Methods. 2014 Mar 9. doi: 10.1038/nmeth.2884.
  PUBMED    24608181
REFERENCE   2  (bases 1 to 6443)
  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"; journalName: "Nat 
            Methods. 2014 Mar 9. doi: 10.1038/nmeth.2884."
COMMENT     Sequence Label: pJT119b
FEATURES             Location/Qualifiers
     source          1..6443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2924..3637)
                     /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"
     RBS             3644..3655
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4610..4696
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(4861..5449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4941..4960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5537..5631)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5652..6311)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6226..6245
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6312..6414)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ctaagctcga ggcaaatggt tatagcgttt taaacagata gtaactgttg ttccctggtc
       61 aactgcactt tttagcaaga tactgccact gtgtaagtca acacatttct tggcaaccat
      121 taaaccgagt cctgttcccg taatatttct gacatttttg ccccgatgaa agggttcaaa
      181 aatttgcttt tggtcctctg gcgaaatgcc aatgccctgg tcggtgactt caaaaataat
      241 ctgttccgaa tctaggctta gggcaatttt aatctgccct cccccgggag agtatttaat
      301 cgcattagat aacagattag ataaaataga ccgcactaac ctttcatcca ccaatgcctt
      361 cgtatcttga gcgctacaaa taaagtcaaa ataatattgg tcactgacac ttaattgaat
      421 ttcttcgata aattgctgga acaataattt caaatctaac caattaggat taaattccaa
      481 tttgcccgct tcggcacggt taatgattaa aatatcatcc aggagctgta ccatattttt
      541 cacggaattt tgaatacggt gtaagttccg gctacgctta tcgggatcaa gccaggccac
      601 ttcagaattt tccagtaatt gggcagcagc taaggccgta ctgaggggag tacgaaattc
      661 atgggaagcc atggagaaaa aacgagtttt taggcggctt aattcttttt cccgttctaa
      721 agctaggcgg acttcctccg tgcgtcggcg ctcagtgaca tccctttgca cagcaatcca
      781 atgggtgtaa aaacctgttt tattggcaac gggcaccaga ctaaattcca cccaaaactc
      841 actgccatcc ttacgataat taatcacttc aacggtaact gattgccatt gactaatggc
      901 ttgccgcacc ctatctaatt cagtgcgact agtttttggt ccctgtaaaa ctcggggggt
      961 tttgcctagc atttcttcag cagtataacc ggtgatttta gtaaatgctt cattgacata
     1021 gagaattctc ggcccgggat catcaatggg ctccgcttcc gtaatcacca ccgcatcatt
     1081 agtattaacc acaacggatt gtagcaaacg aatttgttct gcctgccgct tttgctcggt
     1141 aatatcggcg acaaccgcca ccaatccggc aatattttct tcactatcca ataggggagc
     1201 agcggaaagg acaatttcaa tccaactacc atcttttttt tgacatttta attctaggct
     1261 aggggaaacc atgcctgata aaactttctg tttaaattgc tgataatctt ccagcaaaat
     1321 gttggatgtt aataattctg gatgggcaat aatttctgtt tctgtccaac caaaaatacg
     1381 ttctgctgta ggattccaaa tctgaatttg gtctgctaaa ttaagggtaa aaatacccct
     1441 gggggaggcc gcaatcaggg attgcagagt atggttagtg tggcggagag cggcttccgt
     1501 tttttgtcgc tcactgattt ccattcttag ggattgattg gtggcggcaa gttgctgggt
     1561 gcgtttttcg actcggtttt ccaaatcttt attgagttgc tgtaattgct catataattc
     1621 cgattgttgg atggcgatcg cgacctgatt ggctagctgt ttcattaact ccacttccca
     1681 cggttgccat ggccgggtaa aagcacattg atgggtaatt aacagccccc aaagataggg
     1741 aaattctgat tcattatcta gggaagaagc acgattatgt tgaagaatgg gcactactaa
     1801 ttttgatttc acgccaaatt gtttgacgaa atcagctagg caaatctcta tgtcatcctg
     1861 gtcaatgtca ttaatggccc gtactttacc tttggtgtag gcttgatggt attcaacggg
     1921 aaaaacttca tcggaaaagg tccgccctaa aatactagga taattggcat tcaccgattc
     1981 cgtaatggcg ctgcccgtgc catcttgcca aatgcgataa attagaactc gatcaacttg
     2041 caacagggtt ttaacttcag cgacggtggt atttaaaacc gtttccaagt ttaaagattg
     2101 gcggatatgt tgggtaattt ggttaataaa tcgttcccgt tgggtttgtt gcttgatgct
     2161 aatttcaatc ctgcggcgtt cttggttttg tctgtgccat aaataacagg tcatggcgat
     2221 cgccagaaaa acaaattcga ttggaattag aaatttgccc atatattagc gaatcccagt
     2281 agaaaaacct tacttaaaat atttttgttt ggcaataatt ttcatgactt ttaacaataa
     2341 agctaaatca tgatccttgg caaaaacgcc gtcaatttgg ggcaggtaac tgcgagcttg
     2401 taccgtgatc tgactgtcgg cacaatccgt aattagtagt agggccggcg gcttagcaga
     2461 caatttttta ataatggtca ctaaatctaa agtagctgaa atatcaattg attcatattt
     2521 aatcccgcaa ccaactagga tcgaatgata ttctccaacg tcaacattgt taaacaaaat
     2581 atcttggaag ttgatagcag aaatttcaaa acttggacat aaaaatcgct tgactgattg
     2641 ataccactgc cgatccattt ctaggatacc aatttttgcc ttcgttttct tgcaggtcag
     2701 ggcagaggct tttttaccgg ctagtttcaa cattggagtc tggcctcaaa tacaaagtct
     2761 gcaaagtctc gaaatattaa accagagaat aaataattaa acgccaaaat atttctagca
     2821 aaaaccccag ttattaaacc gcctaagtcc cccaggaaag ggggatataa cagtatagat
     2881 tttgtcagcc ttcagcttgg ctttaccgtc aaaaaaatca tcatttgtac agttcatcca
     2941 taccatgcgt gatgcccgct gcggttacga actccagcag aaccatatga tcgcgtttct
     3001 cgttcggatc tttagacaga acgctttgcg tgctcagata gtgattgtct ggcagcagaa
     3061 caggaccatc accgattgga gtgttttgct ggtagtgatc agccagctgc acgctgccat
     3121 cctccacgtt gtggcgaatt ttaaaattcg ctttaatgcc atttttttgt ttatcggcgg
     3181 tgatgtaaac attgtggctg ttaaaattgt attccagctt atggcccagg atattgccgt
     3241 cttctttaaa gtcaatgcct ttcagctcaa tgcggtttac cagggtatcg ccttcaaatt
     3301 tcacttccgc acgcgttttg tacgtgccgt catccttaaa ggaaatcgtg cgttcctgca
     3361 catagccttc cggcatggcg gacttgaaga agtcatgctg cttcatatgg tccggataac
     3421 gagcaaagca ctgaacacca taagtcagcg tcgttaccag agtcggccaa ggtaccggca
     3481 gtttaccagt agtacagatg aacttcagcg tcagtttacc attagttgcg tcaccttcac
     3541 cctcgccacg cacggaaaac ttatgaccgt tgacatcacc atccagttcc accagaatag
     3601 ggacgacacc agtgaacagc tcttcgcctt tacgcatcta gtatttctcc tctttagata
     3661 aagttagtaa ttaaacttaa aagttgttta atgtccagcc aataactggt atgtgttggc
     3721 attaaaaatg cgatcctaac aaagtaaaat tgaagaaaag ggggagaaaa tcgtaaagaa
     3781 atcggaaagt tagctgatgc gactaaaact gttacatttc gcaacatgaa ttaacccaga
     3841 cctgttcacc ccttcaggta agctgaggtt gatagagaaa agcacaatgg gctatgagaa
     3901 ttcttttagt ggaggatgat ttgccgctgg cggaaaccct tgctgaagca ttgagtgacc
     3961 agctttacac cgttgatatt gccaccgacg cttccctcgc ctgggactat gcctcccgac
     4021 tggaatatga cctcgttatt ttggatgtga tgctgccgga gttggacggg attaccctct
     4081 gtcaaaaatg gcgatcgcac agttatttaa tgccaatttt gatgatgaca gccagggata
     4141 cgatcaatga taaaatcacg ggcttggatg cgggggcgga tgattatgtg gtcaagccag
     4201 tggatttggg ggagttattt gccagggtgc gagctttgtt gcgtcggggt tgtgcaacgt
     4261 gccaaccagt tttagagtgg gggccaatca ggttggatcc aagcacctat gaagttagtt
     4321 atgacaatga ggttttgtct ttgacccgca aggaatacag cattctggaa ttactactcc
     4381 gcaatggccg tcgggtgcta agtcggagca tgattatcga tagtatctgg aagttggaga
     4441 gtcccccaga ggaagatacg gttaaggtgc atgtgcggag tttgcgacaa aaattaaaaa
     4501 gtgccggttt atcagcagat gccattgaaa cggtccatgg cattgggtat cgtctggcca
     4561 atttaacgga aaaatctttg tgccaaggga aaaactagtc tagaggcatc aaataaaacg
     4621 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     4681 cctgagtagg acaaatccgc cgccctagac ctagggcgtt cggctgcggc gagcggtatc
     4741 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4801 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4861 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4921 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4981 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5041 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5101 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5161 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5221 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5281 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5341 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5401 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5461 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5521 catgactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     5581 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     5641 tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     5701 attctgccga catggaagcc atcacagacg gcatgatgaa cctgaatcgc cagcggcatc
     5761 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
     5821 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
     5881 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     5941 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     6001 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     6061 atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg
     6121 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     6181 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     6241 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     6301 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     6361 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct
     6421 cattttcgcc agatatcgac gtc
//
