LOCUS       sequence-331163-        4124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free mercury sensor with enzymatic output.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4124)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 4124)
  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"
COMMENT     Sequence Label: pJBL7090
FEATURES             Location/Qualifiers
     source          1..4124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(49..153)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     221..239
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(277..299)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     399..418
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     612..634
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     626..643
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     627..643
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1722..1751
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1767..1784
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1840..1857)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2010..2056
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2152..2168)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2152..2168)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2165..2187)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2176..2192
                     /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)."
     promoter        complement(join(2200..2206,2207..2224,2225..2230))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2245..2266
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2383..2400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2553..3141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2633..2652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3312..4103,4104..4124,1..48))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3935..3954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
       61 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
      121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
      181 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      241 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
      301 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      361 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      421 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      481 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
      541 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
      601 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtaagatct cggctgaatt
      661 ctaaagatct atcgcttgac tccgtacatg agtacggaag taaggttacg ctatacgtcg
      721 actctcgagt gagattgttg acggtaccgt attttggatc taggaggaag gatctatgaa
      781 caaaggtgta atgcgaccgg gccatgtgca gctgcgtgta ctggacatga gcaaggccct
      841 ggaacactac gtcgagttgc tgggcctgat cgagatggac cgtgacgacc agggccgtgt
      901 ctatctgaag gcttggaccg aagtggataa gttttccctg gtgctacgcg aggctgacga
      961 gccgggcatg gattttatgg gtttcaaggt tgtggatgag gatgctctcc ggcaactgga
     1021 gcgggatctg atggcatatg gctgtgccgt tgagcagcta cccgcaggtg aactgaacag
     1081 ttgtggccgg cgcgtgcgct tccaggcccc ctccgggcat cacttcgagt tgtatgcaga
     1141 caaggaatat actggaaagt ggggtttgaa tgacgtcaat cccgaggcat ggccgcgcga
     1201 tctgaaaggt atggcggctg tgcgtttcga ccacgccctc atgtatggcg acgaattgcc
     1261 ggcgacctat gacctgttca ccaaggtgct cggtttctat ctggccgaac aggtgctgga
     1321 cgaaaatggc acgcgcgtcg cccagtttct cagtctgtcg accaaggccc acgacgtggc
     1381 cttcattcac catccggaaa aaggccgcct ccatcatgtg tccttccacc tcgaaacctg
     1441 ggaagacttg cttcgcgccg ccgacctgat ctccatgacc gacacatcta tcgatatcgg
     1501 cccaacccgc cacggcctca ctcacggcaa gaccatctac ttcttcgacc cgtccggtaa
     1561 ccgcaacgaa gtgttctgcg ggggagatta caactacccg gaccacaaac cggtgacctg
     1621 gaccaccgac cagctgggca aggcgatctt ttaccacgac cgcattctca acgaacgatt
     1681 catgaccgtg ctgacctgat aaggatctga agcttgggcc cgaacaaaaa ctcatctcag
     1741 aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt taaacggtct
     1801 ccagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     1861 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     1921 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     1981 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2041 gggcctttcg ttttatctgt tgtttgtcgg tgaactctga aaggaggaac tatatccgga
     2101 tcgagatcct ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
     2161 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2221 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2281 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2401 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2461 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaggcc
     2521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3241 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3301 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3361 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3421 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3481 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3541 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3601 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3661 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3721 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3781 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3841 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3901 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3961 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4021 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4081 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaa
//
