LOCUS       sequence-331129-        5882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free cascaded mercury sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5882)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 5882)
  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: pJBL7066
FEATURES             Location/Qualifiers
     source          1..5882
                     /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             788..805
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3483..3512
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3598..3615)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3768..3814
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3910..3926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3910..3926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3923..3945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3934..3950
                     /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(3958..3964,3965..3982,3983..3988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4003..4024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4141..4158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4311..4899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4391..4410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5070..5861,5862..5882,1..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     5693..5712
                     /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 gatctatgag
      781 aggatcgcat caccatcacc atcacggatc caacacgatt aacatcgcta agaacgactt
      841 ctctgacatc gaactggctg ctatcccgtt caacactctg gctgaccatt acggtgagcg
      901 tttagctcgc gaacagttgg cccttgagca tgagtcttac gagatgggtg aagcacgctt
      961 ccgcaagatg tttgagcgtc aacttaaagc tggtgaggtt gcggataacg ctgccgccaa
     1021 gcctctcatc actaccctac tccctaagat gattgcacgc atcaacgact ggtttgagga
     1081 agtgaaagct aagcgcggca agcgcccgac agccttccag ttcctgcaag aaatcaagcc
     1141 ggaagccgta gcgtacatca ccattaagac cactctggct tgcctaacca gtgctgacaa
     1201 tacaaccgtt caggctgtag caagcgcaat cggtcgggcc attgaggacg aggctcgctt
     1261 cggtcgtatc cgtgaccttg aagctaagca cttcaagaaa aacgttgagg aacaactcaa
     1321 caagcgcgta gggcacgtct acaagaaagc atttatgcaa gttgtcgagg ctgacatgct
     1381 ctctaagggt ctactcggtg gcgaggcgtg gtcttcgtgg cataaggaag actctattca
     1441 tgtaggagta cgctgcatcg agatgctcat tgagtcaacc ggaatggtta gcttacaccg
     1501 ccaaaatgct ggcgtagtag gtcaagactc tgagactatc gaactcgcac ctgaatacgc
     1561 tgaggctatc gcaacccgtg caggtgcgct ggctggcatc tctccgatgt tccaaccttg
     1621 cgtagttcct cctaagccgt ggactggcat tactggtggt ggctattggg ctaacggtcg
     1681 tcgtcctctg gcgctggtgc gtactcacag taagaaagca ctgatgcgct acgaagacgt
     1741 ttacatgcct gaggtgtaca aagcgattaa cattgcgcaa aacaccgcat ggaaaatcaa
     1801 caagaaagtc ctagcggtcg ccaacgtaat caccaagtgg aagcattgtc cggtcgagga
     1861 catccctgcg attgagcgtg aagaactccc gatgaaaccg gaagacatcg acatgaatcc
     1921 tgaggctctc accgcgtgga aacgtgctgc cgctgctgtg taccgcaagg acaaggctcg
     1981 caagtctcgc cgtatcagcc ttgagttcat gcttgagcaa gccaataagt ttgctaacca
     2041 taaggccatc tggttccctt acaacatgga ctggcgcggt cgtgtttacg ctgtgtcaat
     2101 gttcaacccg caaggtaacg atatgaccaa aggactgctt acgctggcga aaggtaaacc
     2161 aatcggtaag gaaggttact actggctgaa aatccacggt gcaaactgtg cgggtgtcga
     2221 taaggttccg ttccctgagc gcatcaagtt cattgaggaa aaccacgaga acatcatggc
     2281 ttgcgctaag tctccactgg agaacacttg gtgggctgag caagattctc cgttctgctt
     2341 ccttgcattc tgctttgagt acgctggggt acagcaccac ggcctgagct ataactgctc
     2401 ccttccgctg gcgtttgacg ggtcttgctc tggcatccag cacttctccg cgatgctccg
     2461 agatgaggta ggtggtcgcg cggttaactt gcttcctagt gaaaccgttc aggacatcta
     2521 cgggattgtt gctaagaaag tcaacgagat tctacaagca gacgcaatca atgggaccga
     2581 taacgaagta gttaccgtga ccgatgagaa cactggtgaa atctctgaga aagtcaagct
     2641 gggcactaag gcactggctg gtcaatggct ggcttacggt gttactcgca gtgtgactaa
     2701 gcgttcagtc atgacgctgg cttacgggtc caaagagttc ggcttccgtc aacaagtgct
     2761 ggaagatacc attcagccag ctattgattc cggcaagggt ctgatgttca ctcagccgaa
     2821 tcaggctgct ggatacatgg ctaagctgat ttgggaatct gtgagcgtga cggtggtagc
     2881 tgcggttgaa gcaatgaact ggcttaagtc tgctgctaag ctgctggctg ctgaggtcaa
     2941 agataagaag actggagaga ttcttcgcaa gcgttgcgct gcgcattggg taactcctga
     3001 tggtttccct gtgtggcagg aatacaagaa gcctattaaa acgcgcatct ctataatgtt
     3061 cctcggtcag ttcaccttaa agcctaccat taacaccaac aaagatagcg agattgatgc
     3121 acgcaaacag gtgtctggta tcgctcctaa ctttgtacac agccaagacg gtagccacct
     3181 tcgtaagact gtagtgtggg cacacgagaa gtacggaatc gaatcttttg cactgattca
     3241 cgactccttc ggtaccattc cggctgacgc tgcgaacctg ttcaaagcag tgcgcgaaac
     3301 tatggttgac acatatgagt cttgtgatgt actggctgat ttctacgacc agttcgctga
     3361 ccagttgcac gagtctcaat tggacaaaat gccagcactt ccggctaaag gtaacttgaa
     3421 cctccgtgac atcttagagt cggacttcgc gttcgcgtaa taaggatctg aagcttgggc
     3481 ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc
     3541 attgagttta aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct
     3601 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     3661 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     3721 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     3781 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aactctgaaa
     3841 ggaggaacta tatccggatc gagatcctct agagtcgacc tgcaggcatg caagcttggc
     3901 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3961 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4021 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4081 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4141 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4201 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4261 aaaaggccag caaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4321 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4381 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4441 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4501 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4561 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4621 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4681 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4741 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4801 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4861 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4921 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4981 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5041 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5101 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5161 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5221 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5281 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5341 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5401 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5461 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5521 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5581 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5641 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5701 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5761 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5821 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5881 aa
//
