LOCUS       sequence-324796-        2518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free lac sensor with RFP output.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2518)
  AUTHORS   Jung JK, Rasor BJ, Rybnicky GA, Silverman AD, Standeven J, Kuhn R, 
            Granito T, Ekas HM, Wang BM, Karim AS, Lucks JB, Jewett MC
  TITLE     At-Home, Cell-Free Synthetic Biology Education Modules for 
            Transcriptional Regulation and Environmental Water Quality 
            Monitoring.
  JOURNAL   ACS Synth Biol. 2023 Oct 20;12(10):2909-2921. doi: 
            10.1021/acssynbio.3c00223. Epub 2023 Sep 12.
  PUBMED    37699423
REFERENCE   2  (bases 1 to 2518)
  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/acssynbio.3c00223"; journalName: "ACS Synth Biol"; date: 
            "2023-10-20- 20"; volume: "12"; issue: "10"; pages: "2909-2921"
COMMENT     Sequence Label: pJBL7080
FEATURES             Location/Qualifiers
     source          1..2518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     194..213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(343..348,349..366,367..373)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    381..397
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     386..408
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     RBS             448..470
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             478..1155
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1207..1225)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1221..1268
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1286..2101)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2009..2028
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2093..2112
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2223..2518,1..293)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
       61 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      121 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      181 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      241 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      301 gcaacgcgat cccgcgaaat tcactcatta ggcaccccag gctttacact ttatgcttcc
      361 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctgaga
      421 ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
      481 gcttcctccg aagacgttat caaagagttc atgcgtttca aagttcgtat ggaaggttcc
      541 gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc
      601 cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg
      661 tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga catcccggac
      721 tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac
      781 ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt catctacaaa
      841 gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg
      901 ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc
      961 aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa aaccacctac
     1021 atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa actggacatc
     1081 acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga aggtcgtcac
     1141 tccaccggtg cttaagtcga ccggctgcta acaaagcccg aaaggaagct gagttggctg
     1201 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1261 gttttttgct gaaagccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     1321 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1381 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     1441 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1501 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
     1561 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     1621 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     1681 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     1741 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     1801 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     1861 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     1921 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     1981 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     2041 catccatgtt ggaatttaat cgcggcttcg agcaagacgt ttcccgttga atatggctca
     2101 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     2161 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtgagat caaaggatct
     2221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2341 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccgg
//
