LOCUS       sequence-331168-        3933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free arsenic sensing reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3933)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 3933)
  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: pJBL7049
FEATURES             Location/Qualifiers
     source          1..3933
                     /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             795..1508
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITTDKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(825..853)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1448..1468
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1531..1560
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1576..1593
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1649..1666)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1819..1865
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1961..1977)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1961..1977)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1974..1996)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1985..2001
                     /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(2009..2015,2016..2033,2034..2039))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2054..2075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2192..2209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2362..2950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2442..2461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3121..3912,3913..3933,1..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     3744..3763
                     /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 atctatgagc aaaggagaag aacttttcac tggagttgtc ccaattcttg
      841 ttgaattaga tggtgatgtt aatgggcaca aattttctgt ccgtggagag ggtgaaggtg
      901 atgctacaaa cggaaaactc acccttaaat ttatttgcac tactggaaaa ctacctgttc
      961 cgtggccaac acttgtcact actctgacct atggtgttca atgcttttcc cgttatccgg
     1021 atcacatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaac
     1081 gcactatatc tttcaaagat gacgggacct acaagacgcg tgctgaagtc aagtttgaag
     1141 gtgataccct tgttaatcgt atcgagttaa agggtattga ttttaaagaa gatggaaaca
     1201 ttcttggaca caaactcgag tacaacttta actcacacaa tgtatacatc acgacagaca
     1261 aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacgttgaa gatggttccg
     1321 ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
     1381 cagacaacca ttacctgtcg acacaatctg tcctttcgaa agatcccaac gaaaagcgtg
     1441 accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc atggatgagc
     1501 tctacaaata aggatctgaa gcttgggccc gaacaaaaac tcatctcaga agaggatctg
     1561 aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacggtctc cagcttggct
     1621 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     1681 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     1741 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     1801 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1861 tttatctgtt gtttgtcggt gaactctgaa aggaggaact atatccggat cgagatcctc
     1921 tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1981 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     2041 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     2101 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2161 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2221 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2281 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaggcca ggaaccgtaa
     2341 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2401 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2461 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2521 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2581 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2641 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccagtaagac acgacttatc
     2701 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2761 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2821 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2881 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2941 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3001 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3061 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3121 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3181 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3241 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3301 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3361 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3421 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3481 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3541 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3601 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3661 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3721 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3781 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3841 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3901 cgtttctggg tgagcaaaaa caggaaggca aaa
//
