LOCUS       sequence-171059-        4091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A weak promoter drives the expression of a hypersensitive luxR 
            mutant, which regulates expression of GFP under the control of a 
            strong RBS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4091)
  AUTHORS   Magaraci MS, Bermudez JG, Yogish D, Pak DH, Mollov V, Tycko J, 
            Issadore D, Mannickarottu SG, Chow BY
  TITLE     Toolbox for Exploring Modular Gene Regulation in Synthetic Biology 
            Training.
  JOURNAL   ACS Synth Biol. 2016 Apr 25.
  PUBMED    27111289
REFERENCE   2  (bases 1 to 4091)
  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: "ACS Synth 
            Biol. 2016 Apr 25."
COMMENT     Sequence Label: BC-A1-014
FEATURES             Location/Qualifiers
     source          1..4091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..714
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(31..59)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     654..674
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      768..839
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      855..882
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      904..961
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(985..1002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1156..1744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1236..1255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1927..2021)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2042..2701)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2616..2635
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2863..2881
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2885..2928)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        2938..2972
                     /label=BBa_J23109
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.04"
     RBS             2979..2990
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3802..3873
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3889..3916
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             4074..4085
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgcgtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
       61 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      121 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      181 gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca catgaaacag
      241 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
      301 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
      361 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
      421 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
      481 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
      541 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
      601 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
      661 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaataataa
      721 tctagacact gatagtgcta gtgtagatca ctactagagc caggcatcaa ataaaacgaa
      781 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
      841 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatacc cgggtactag
      901 aggtccggca aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag
      961 attacttcgc gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1141 cgcgttgctg gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg
     1201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaagtatc tcagttcggt
     1381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1561 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1741 tcaagaagat cctttgatct tttctacggg gtcagacgct cagtggaacg aaaactcacg
     1801 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1861 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagctcgag
     1921 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
     1981 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa
     2041 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     2101 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     2161 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     2221 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     2281 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     2341 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     2401 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     2461 caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt
     2521 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     2581 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     2641 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     2701 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     2761 ttatttcatt atggtgaaag ttggaacctc ttacgtgccc gatcaactcg agtgccacct
     2821 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2881 cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg gctgcagttt
     2941 acagctagct cagtcctagg gactgtgcta gcggatccaa agaggagaaa gaattcatga
     3001 aaaacataaa tgccgacgac acatacagaa taattaataa aattaaagcg tgtagaagca
     3061 ataatgatat taatcaatgc ttatctgata tggctaaaat ggtacattgt gaatattatt
     3121 tactcgcgtt catttatcct cattctatgg ttaaatcaga tatttcaatc ctagataatt
     3181 accctaaaaa atggaggcaa tattatgatg acgctaattt aataaaatat gatcctatag
     3241 tagattattc taactccaat cattcaccaa ttaattggaa tatatttgaa aacaatgctg
     3301 taaataaaaa atctccaaat gtaattaaag aagcgaaaac agcaggtctt atcactgggt
     3361 ttagtttccc tattcatacg gctaacaatg gcttcggaat gcttagtttt gcacattcag
     3421 aaaaagacaa ctatatagat agtttatttt tacatgcgtg tatgaacata ccattaattg
     3481 ttccttctct agttgataat tatcgaaaaa taaatatagc aaataataaa tcaaacaacg
     3541 atttaaccaa aagagaaaaa gaatgtttag cgtgggcatg cgaaggaaaa agctcttggg
     3601 atatttcaaa aatattaggt tgcagtgagc gtactgtcac tttccattta accaatgcgc
     3661 aaatgaaact caatacaaca aaccgctgcc aaagtatttc taaagcaatt ttaacaggag
     3721 caattgattg cccatacttt aaaaattaat aagcggccgc cactgatagt gctagtgtag
     3781 atcactacta gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     3841 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     3901 ggtgggcctt tctgcgttta taggtaccta ctagagaggt tctgttaagt aactgaaccc
     3961 aatgtcgtta gtgacgctta cctcttaaga ggtcactgac ctaacaaagc ttacctgtag
     4021 gatcgtacag gtttacgcaa gaaaatggtt tgttatagtc gaataaaact agtaaagagg
     4081 agaaatagca t
//
