LOCUS       sequence-331125-        5880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free cascaded cadmium sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5880)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 5880)
  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: pJBL7064
FEATURES             Location/Qualifiers
     source          1..5880
                     /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             786..803
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3596..3613)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3766..3812
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3908..3924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3908..3924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3921..3943)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3932..3948
                     /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(3956..3962,3963..3980,3981..3986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4001..4022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4139..4156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4309..4897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4389..4408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5068..5859,5860..5880,1..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     5691..5710
                     /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 ctaaaagatc tataacttga ctctgtagtt gctacagggt gtgcaatcgg ttacgtcgac
      721 tctcgagtga gattgttgac ggtaccgtat tttggatcta ggaggaagga tctatgagag
      781 gatcgcatca ccatcaccat cacggatcca acacgattaa catcgctaag aacgacttct
      841 ctgacatcga actggctgct atcccgttca acactctggc tgaccattac ggtgagcgtt
      901 tagctcgcga acagttggcc cttgagcatg agtcttacga gatgggtgaa gcacgcttcc
      961 gcaagatgtt tgagcgtcaa cttaaagctg gtgaggttgc ggataacgct gccgccaagc
     1021 ctctcatcac taccctactc cctaagatga ttgcacgcat caacgactgg tttgaggaag
     1081 tgaaagctaa gcgcggcaag cgcccgacag ccttccagtt cctgcaagaa atcaagccgg
     1141 aagccgtagc gtacatcacc attaagacca ctctggcttg cctaaccagt gctgacaata
     1201 caaccgttca ggctgtagca agcgcaatcg gtcgggccat tgaggacgag gctcgcttcg
     1261 gtcgtatccg tgaccttgaa gctaagcact tcaagaaaaa cgttgaggaa caactcaaca
     1321 agcgcgtagg gcacgtctac aagaaagcat ttatgcaagt tgtcgaggct gacatgctct
     1381 ctaagggtct actcggtggc gaggcgtggt cttcgtggca taaggaagac tctattcatg
     1441 taggagtacg ctgcatcgag atgctcattg agtcaaccgg aatggttagc ttacaccgcc
     1501 aaaatgctgg cgtagtaggt caagactctg agactatcga actcgcacct gaatacgctg
     1561 aggctatcgc aacccgtgca ggtgcgctgg ctggcatctc tccgatgttc caaccttgcg
     1621 tagttcctcc taagccgtgg actggcatta ctggtggtgg ctattgggct aacggtcgtc
     1681 gtcctctggc gctggtgcgt actcacagta agaaagcact gatgcgctac gaagacgttt
     1741 acatgcctga ggtgtacaaa gcgattaaca ttgcgcaaaa caccgcatgg aaaatcaaca
     1801 agaaagtcct agcggtcgcc aacgtaatca ccaagtggaa gcattgtccg gtcgaggaca
     1861 tccctgcgat tgagcgtgaa gaactcccga tgaaaccgga agacatcgac atgaatcctg
     1921 aggctctcac cgcgtggaaa cgtgctgccg ctgctgtgta ccgcaaggac aaggctcgca
     1981 agtctcgccg tatcagcctt gagttcatgc ttgagcaagc caataagttt gctaaccata
     2041 aggccatctg gttcccttac aacatggact ggcgcggtcg tgtttacgct gtgtcaatgt
     2101 tcaacccgca aggtaacgat atgaccaaag gactgcttac gctggcgaaa ggtaaaccaa
     2161 tcggtaagga aggttactac tggctgaaaa tccacggtgc aaactgtgcg ggtgtcgata
     2221 aggttccgtt ccctgagcgc atcaagttca ttgaggaaaa ccacgagaac atcatggctt
     2281 gcgctaagtc tccactggag aacacttggt gggctgagca agattctccg ttctgcttcc
     2341 ttgcattctg ctttgagtac gctggggtac agcaccacgg cctgagctat aactgctccc
     2401 ttccgctggc gtttgacggg tcttgctctg gcatccagca cttctccgcg atgctccgag
     2461 atgaggtagg tggtcgcgcg gttaacttgc ttcctagtga aaccgttcag gacatctacg
     2521 ggattgttgc taagaaagtc aacgagattc tacaagcaga cgcaatcaat gggaccgata
     2581 acgaagtagt taccgtgacc gatgagaaca ctggtgaaat ctctgagaaa gtcaagctgg
     2641 gcactaaggc actggctggt caatggctgg cttacggtgt tactcgcagt gtgactaagc
     2701 gttcagtcat gacgctggct tacgggtcca aagagttcgg cttccgtcaa caagtgctgg
     2761 aagataccat tcagccagct attgattccg gcaagggtct gatgttcact cagccgaatc
     2821 aggctgctgg atacatggct aagctgattt gggaatctgt gagcgtgacg gtggtagctg
     2881 cggttgaagc aatgaactgg cttaagtctg ctgctaagct gctggctgct gaggtcaaag
     2941 ataagaagac tggagagatt cttcgcaagc gttgcgctgc gcattgggta actcctgatg
     3001 gtttccctgt gtggcaggaa tacaagaagc ctattaaaac gcgcatctct ataatgttcc
     3061 tcggtcagtt caccttaaag cctaccatta acaccaacaa agatagcgag attgatgcac
     3121 gcaaacaggt gtctggtatc gctcctaact ttgtacacag ccaagacggt agccaccttc
     3181 gtaagactgt agtgtgggca cacgagaagt acggaatcga atcttttgca ctgattcacg
     3241 actccttcgg taccattccg gctgacgctg cgaacctgtt caaagcagtg cgcgaaacta
     3301 tggttgacac atatgagtct tgtgatgtac tggctgattt ctacgaccag ttcgctgacc
     3361 agttgcacga gtctcaattg gacaaaatgc cagcacttcc ggctaaaggt aacttgaacc
     3421 tccgtgacat cttagagtcg gacttcgcgt tcgcgtaata aggatctgaa gcttgggccc
     3481 gaaaaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat
     3541 tgagtttaaa cggtctccag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     3601 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     3661 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     3721 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     3781 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa ctctgaaagg
     3841 aggaactata tccggatcga gatcctctag agtcgacctg caggcatgca agcttggcgt
     3901 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3961 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4021 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4081 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4141 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4201 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4261 aaggccagca aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
//
