LOCUS       sequence-324804-        4144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free arsenic sensor with enzymatic output.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4144)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 4144)
  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: pJBL7091
FEATURES             Location/Qualifiers
     source          1..4144
                     /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             1741..1770
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1786..1803
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1859..1876)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2029..2075
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2171..2187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2171..2187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2184..2206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2195..2211
                     /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(2219..2225,2226..2243,2244..2249))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2264..2285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2402..2419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2573..3161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2653..2672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3332..4123,4124..4144,1..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     3955..3974
                     /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 acacattcgt taagtcatat atgtttttga cttatccgct tcgaagagat
      721 ataatacctg caaacgtcga ctctcgagtg agattgttga cggtaccgta ttttggatct
      781 aggaggaagg atctatgaac aaaggtgtaa tgcgaccggg ccatgtgcag ctgcgtgtac
      841 tggacatgag caaggccctg gaacactacg tcgagttgct gggcctgatc gagatggacc
      901 gtgacgacca gggccgtgtc tatctgaagg cttggaccga agtggataag ttttccctgg
      961 tgctacgcga ggctgacgag ccgggcatgg attttatggg tttcaaggtt gtggatgagg
     1021 atgctctccg gcaactggag cgggatctga tggcatatgg ctgtgccgtt gagcagctac
     1081 ccgcaggtga actgaacagt tgtggccggc gcgtgcgctt ccaggccccc tccgggcatc
     1141 acttcgagtt gtatgcagac aaggaatata ctggaaagtg gggtttgaat gacgtcaatc
     1201 ccgaggcatg gccgcgcgat ctgaaaggta tggcggctgt gcgtttcgac cacgccctca
     1261 tgtatggcga cgaattgccg gcgacctatg acctgttcac caaggtgctc ggtttctatc
     1321 tggccgaaca ggtgctggac gaaaatggca cgcgcgtcgc ccagtttctc agtctgtcga
     1381 ccaaggccca cgacgtggcc ttcattcacc atccggaaaa aggccgcctc catcatgtgt
     1441 ccttccacct cgaaacctgg gaagacttgc ttcgcgccgc cgacctgatc tccatgaccg
     1501 acacatctat cgatatcggc ccaacccgcc acggcctcac tcacggcaag accatctact
     1561 tcttcgaccc gtccggtaac cgcaacgaag tgttctgcgg gggagattac aactacccgg
     1621 accacaaacc ggtgacctgg accaccgacc agctgggcaa ggcgatcttt taccacgacc
     1681 gcattctcaa cgaacgattc atgaccgtgc tgacctgata aggatctgaa gcttgggccc
     1741 gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat
     1801 cattgagttt aaacggtctc cagcttggct gttttggcgg atgagagaag attttcagcc
     1861 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     1921 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     1981 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     2041 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaactctgaa
     2101 aggaggaact atatccggat cgagatcctc tagagtcgac ctgcaggcat gcaagcttgg
     2161 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2221 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2281 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2341 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2401 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2461 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2521 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2581 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2641 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2701 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2761 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2821 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2881 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2941 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3001 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3061 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3121 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3181 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3241 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3301 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3361 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3421 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3481 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3541 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3601 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3661 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3721 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3781 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3841 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3901 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3961 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4021 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4081 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4141 aaaa
//
