LOCUS       sequence-331124-        5878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell-free cascaded copper sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5878)
  AUTHORS   Jung JK, Rasor BJ, Rybnicky GA, Silverman AD, Standeven J, Kuhn R, 
            Granito T, Ekas HM, Wang BM, Karim AS, Lucks JB, Jewett MC
  TITLE     At-Home, Cell-Free Synthetic Biology Education Modules for 
            Transcriptional Regulation and Environmental Water Quality 
            Monitoring.
  JOURNAL   ACS Synth Biol. 2023 Oct 20;12(10):2909-2921. doi: 
            10.1021/acssynbio.3c00223. Epub 2023 Sep 12.
  PUBMED    37699423
REFERENCE   2  (bases 1 to 5878)
  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"; doi: 
            "10.1021/acssynbio.3c00223"; journalName: "ACS Synth Biol"; date: 
            "2023-10-20- 20"; volume: "12"; issue: "10"; pages: "2909-2921"
COMMENT     Sequence Label: pJBL7062
FEATURES             Location/Qualifiers
     source          1..5878
                     /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             784..801
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3479..3508
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3594..3611)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3764..3810
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3906..3922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3906..3922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3919..3941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3930..3946
                     /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(3954..3960,3961..3978,3979..3984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3999..4020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4137..4154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4307..4895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4387..4406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5066..5857,5858..5878,1..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     5689..5708
                     /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 tttcttgacc ttccccttgc tggaaggttt atcctcggtt acgtcgactc
      721 tcgagtgaga ttgttgacgg taccgtattt tggatctaga aggaaggatc tatgagagga
      781 tcgcatcacc atcaccatca cggatccaac acgattaaca tcgctaagaa cgacttctct
      841 gacatcgaac tggctgctat cccgttcaac actctggctg accattacgg tgagcgttta
      901 gctcgcgaac agttggccct tgagcatgag tcttacgaga tgggtgaagc acgcttccgc
      961 aagatgtttg agcgtcaact taaagctggt gaggttgcgg ataacgctgc cgccaagcct
     1021 ctcatcacta ccctactccc taagatgatt gcacgcatca acgactggtt tgaggaagtg
     1081 aaagctaagc gcggcaagcg cccgacagcc ttccagttcc tgcaagaaat caagccggaa
     1141 gccgtagcgt acatcaccat taagaccact ctggcttgcc taaccagtgc tgacaataca
     1201 accgttcagg ctgtagcaag cgcaatcggt cgggccattg aggacgaggc tcgcttcggt
     1261 cgtatccgtg accttgaagc taagcacttc aagaaaaacg ttgaggaaca actcaacaag
     1321 cgcgtagggc acgtctacaa gaaagcattt atgcaagttg tcgaggctga catgctctct
     1381 aagggtctac tcggtggcga ggcgtggtct tcgtggcata aggaagactc tattcatgta
     1441 ggagtacgct gcatcgagat gctcattgag tcaaccggaa tggttagctt acaccgccaa
     1501 aatgctggcg tagtaggtca agactctgag actatcgaac tcgcacctga atacgctgag
     1561 gctatcgcaa cccgtgcagg tgcgctggct ggcatctctc cgatgttcca accttgcgta
     1621 gttcctccta agccgtggac tggcattact ggtggtggct attgggctaa cggtcgtcgt
     1681 cctctggcgc tggtgcgtac tcacagtaag aaagcactga tgcgctacga agacgtttac
     1741 atgcctgagg tgtacaaagc gattaacatt gcgcaaaaca ccgcatggaa aatcaacaag
     1801 aaagtcctag cggtcgccaa cgtaatcacc aagtggaagc attgtccggt cgaggacatc
     1861 cctgcgattg agcgtgaaga actcccgatg aaaccggaag acatcgacat gaatcctgag
     1921 gctctcaccg cgtggaaacg tgctgccgct gctgtgtacc gcaaggacaa ggctcgcaag
     1981 tctcgccgta tcagccttga gttcatgctt gagcaagcca ataagtttgc taaccataag
     2041 gccatctggt tcccttacaa catggactgg cgcggtcgtg tttacgctgt gtcaatgttc
     2101 aacccgcaag gtaacgatat gaccaaagga ctgcttacgc tggcgaaagg taaaccaatc
     2161 ggtaaggaag gttactactg gctgaaaatc cacggtgcaa actgtgcggg tgtcgataag
     2221 gttccgttcc ctgagcgcat caagttcatt gaggaaaacc acgagaacat catggcttgc
     2281 gctaagtctc cactggagaa cacttggtgg gctgagcaag attctccgtt ctgcttcctt
     2341 gcattctgct ttgagtacgc tggggtacag caccacggcc tgagctataa ctgctccctt
     2401 ccgctggcgt ttgacgggtc ttgctctggc atccagcact tctccgcgat gctccgagat
     2461 gaggtaggtg gtcgcgcggt taacttgctt cctagtgaaa ccgttcagga catctacggg
     2521 attgttgcta agaaagtcaa cgagattcta caagcagacg caatcaatgg gaccgataac
     2581 gaagtagtta ccgtgaccga tgagaacact ggtgaaatct ctgagaaagt caagctgggc
     2641 actaaggcac tggctggtca atggctggct tacggtgtta ctcgcagtgt gactaagcgt
     2701 tcagtcatga cgctggctta cgggtccaaa gagttcggct tccgtcaaca agtgctggaa
     2761 gataccattc agccagctat tgattccggc aagggtctga tgttcactca gccgaatcag
     2821 gctgctggat acatggctaa gctgatttgg gaatctgtga gcgtgacggt ggtagctgcg
     2881 gttgaagcaa tgaactggct taagtctgct gctaagctgc tggctgctga ggtcaaagat
     2941 aagaagactg gagagattct tcgcaagcgt tgcgctgcgc attgggtaac tcctgatggt
     3001 ttccctgtgt ggcaggaata caagaagcct attaaaacgc gcatctctat aatgttcctc
     3061 ggtcagttca ccttaaagcc taccattaac accaacaaag atagcgagat tgatgcacgc
     3121 aaacaggtgt ctggtatcgc tcctaacttt gtacacagcc aagacggtag ccaccttcgt
     3181 aagactgtag tgtgggcaca cgagaagtac ggaatcgaat cttttgcact gattcacgac
     3241 tccttcggta ccattccggc tgacgctgcg aacctgttca aagcagtgcg cgaaactatg
     3301 gttgacacat atgagtcttg tgatgtactg gctgatttct acgaccagtt cgctgaccag
     3361 ttgcacgagt ctcaattgga caaaatgcca gcacttccgg ctaaaggtaa cttgaacctc
     3421 cgtgacatct tagagtcgga cttcgcgttc gcgtaataag gatctgaagc ttgggcccga
     3481 acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcattg
     3541 agtttaaacg gtctccagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     3601 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     3661 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     3721 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     3781 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaact ctgaaaggag
     3841 gaactatatc cggatcgaga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa
     3901 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3961 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4021 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4081 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4141 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4261 ggccagcaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4321 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4381 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4441 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4501 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4561 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4621 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4681 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4741 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4801 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaa
//
