LOCUS       sequence-331164-        4143 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 4143)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 4143)
  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..4143
                     /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(2572..3160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2652..2671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3331..4122,4123..4143,1..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     3954..3973
                     /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 gcaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2581 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2641 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2701 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2761 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2821 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2881 tgagtccaac ccagtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2941 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3001 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3061 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3121 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3181 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3241 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3301 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3361 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3421 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3481 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3541 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3601 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3661 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3721 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3781 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3841 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3901 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3961 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4021 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4081 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4141 aaa
//
