LOCUS       sequence-366447-        2767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free lac sensor with XylE output.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2767)
  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 2767)
  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: pJBL7082
FEATURES             Location/Qualifiers
     source          1..2767
                     /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)"
     primer_bind     complement(1456..1474)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1470..1517
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1535..2350)
                     /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     2258..2277
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2342..2361
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2472..2767,1..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 aacaaaggtg taatgcgacc gggccatgtg cagctgcgtg tactggacat gagcaaggcc
      541 ctggaacact acgtcgagtt gctgggcctg atcgagatgg accgtgacga ccagggccgt
      601 gtctatctga aggcttggac cgaagtggat aagttttccc tggtgctacg cgaggctgac
      661 gagccgggca tggattttat gggtttcaag gttgtggatg aggatgctct ccggcaactg
      721 gagcgggatc tgatggcata tggctgtgcc gttgagcagc tacccgcagg tgaactgaac
      781 agttgtggcc ggcgcgtgcg cttccaggcc ccctccgggc atcacttcga gttgtatgca
      841 gacaaggaat atactggaaa gtggggtttg aatgacgtca atcccgaggc atggccgcgc
      901 gatctgaaag gtatggcggc tgtgcgtttc gaccacgccc tcatgtatgg cgacgaattg
      961 ccggcgacct atgacctgtt caccaaggtg ctcggtttct atctggccga acaggtgctg
     1021 gacgaaaatg gcacgcgcgt cgcccagttt ctcagtctgt cgaccaaggc ccacgacgtg
     1081 gccttcattc accatccgga aaaaggccgc ctccatcatg tgtccttcca cctcgaaacc
     1141 tgggaagact tgcttcgcgc cgccgacctg atctccatga ccgacacatc tatcgatatc
     1201 ggcccaaccc gccacggcct cactcacggc aagaccatct acttcttcga cccgtccggt
     1261 aaccgcaacg aagtgttctg cgggggagat tacaactacc cggaccacaa accggtgacc
     1321 tggaccaccg accagctggg caaggcgatc ttttaccacg accgcattct caacgaacga
     1381 ttcatgaccg tgctgaccta agtcgtcgac cggctgctaa caaagcccga aaggaagctg
     1441 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1501 tcttgagggg ttttttgctg aaagccaatt ctgattagaa aaactcatcg agcatcaaat
     1561 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     1621 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     1681 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     1741 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct
     1801 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     1861 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     1921 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     1981 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     2041 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     2101 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     2161 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     2221 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     2281 ataaatcagc atccatgttg gaatttaatc gcggcttcga gcaagacgtt tcccgttgaa
     2341 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     2401 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtgagatc
     2461 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2521 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2581 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2641 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2701 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2761 ttaccgg
//
