LOCUS       sequence-324797-        1855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free lac sensor with Mango output.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1855)
  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 1855)
  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: pJBL7081
FEATURES             Location/Qualifiers
     source          1..1855
                     /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"
     terminator      558..605
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(623..1438)
                     /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     1346..1365
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1430..1449
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(1560..1855,1..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 acagctggca
      421 cgtacgaagg aaggattggt atgtggtata ttcgtacgtg ccgccggcca tggtcccagc
      481 ctcctcgctg gcggccggtg ggcaacattc cgaggggacc gtcccctcgg taatggcgaa
      541 tgggacccat cgtagatcta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      601 ttttgctgaa agccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
      661 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
      721 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
      781 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
      841 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
      901 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
      961 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     1021 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     1081 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     1141 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     1201 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     1261 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     1321 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     1381 ccatgttgga atttaatcgc ggcttcgagc aagacgtttc ccgttgaata tggctcataa
     1441 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     1501 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtgagatcaa aggatcttct
     1561 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1621 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1681 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1741 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1801 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accgg
//
