LOCUS       sequence-331178-        4116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free lead sensor with XylE output.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4116)
  TITLE     Sensitizing cell-free molecular detection using cascaded genetic 
            circuitry
REFERENCE   2  (bases 1 to 4116)
  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: pJBL7093
FEATURES             Location/Qualifiers
     source          1..4116
                     /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"
     primer_bind     complement(1832..1849)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2002..2048
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2144..2160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2144..2160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2157..2179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2168..2184
                     /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(2192..2198,2199..2216,2217..2222))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2237..2258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2375..2392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2545..3133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2625..2644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3304..4095,4096..4116,1..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     3927..3946
                     /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 tatgtcttga ctctatagta actagagggt gttaaatcgg caacgtcgac
      721 tctcgagtga gattgttgac ggtaccgtat tttggatcta ggaggaagga tctatgaaca
      781 aaggtgtaat gcgaccgggc catgtgcagc tgcgtgtact ggacatgagc aaggccctgg
      841 aacactacgt cgagttgctg ggcctgatcg agatggaccg tgacgaccag ggccgtgtct
      901 atctgaaggc ttggaccgaa gtggataagt tttccctggt gctacgcgag gctgacgagc
      961 cgggcatgga ttttatgggt ttcaaggttg tggatgagga tgctctccgg caactggagc
     1021 gggatctgat ggcatatggc tgtgccgttg agcagctacc cgcaggtgaa ctgaacagtt
     1081 gtggccggcg cgtgcgcttc caggccccct ccgggcatca cttcgagttg tatgcagaca
     1141 aggaatatac tggaaagtgg ggtttgaatg acgtcaatcc cgaggcatgg ccgcgcgatc
     1201 tgaaaggtat ggcggctgtg cgtttcgacc acgccctcat gtatggcgac gaattgccgg
     1261 cgacctatga cctgttcacc aaggtgctcg gtttctatct ggccgaacag gtgctggacg
     1321 aaaatggcac gcgcgtcgcc cagtttctca gtctgtcgac caaggcccac gacgtggcct
     1381 tcattcacca tccggaaaaa ggccgcctcc atcatgtgtc cttccacctc gaaacctggg
     1441 aagacttgct tcgcgccgcc gacctgatct ccatgaccga cacatctatc gatatcggcc
     1501 caacccgcca cggcctcact cacggcaaga ccatctactt cttcgacccg tccggtaacc
     1561 gcaacgaagt gttctgcggg ggagattaca actacccgga ccacaaaccg gtgacctgga
     1621 ccaccgacca gctgggcaag gcgatctttt accacgaccg cattctcaac gaacgattca
     1681 tgaccgtgct gacctgagga tctgaagctt gggcccgaaa aaaaactcat ctcagaagag
     1741 gatctgaata gcgccgtcga ccatcatcat catcattgag tttaaacggt ctccagcttg
     1801 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
     1861 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     1921 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     1981 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2041 cgttttatct gttgtttgtc ggtgaactct gaaaggagga actatatccg gatcgagatc
     2101 ctctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
     2161 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2221 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2281 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2341 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2401 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2461 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaagg ccaggaaccg
     2521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3301 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3361 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3421 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3481 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3541 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3601 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3661 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3721 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3781 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3841 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3901 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3961 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4021 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4081 cagcgtttct gggtgagcaa aaacaggaag gcaaaa
//
