LOCUS       sequence-297839-        2921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the tetO sequence 2BP dowsntream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2921)
  AUTHORS   Jung JK, Alam KK, Verosloff MS, Capdevila DA, Desmau M, Clauer PR, 
            Lee JW, Nguyen PQ, Pasten PA, Matiasek SJ, Gaillard JF, Giedroc DP, 
            Collins JJ, Lucks JB
  TITLE     Cell-free biosensors for rapid detection of water contaminants.
  JOURNAL   Nat Biotechnol. 2020 Jul 6. pii: 10.1038/s41587-020-0571-7. doi: 
            10.1038/s41587-020-0571-7.
  PUBMED    32632301
REFERENCE   2  (bases 1 to 2921)
  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"; journalName: "Nat 
            Biotechnol. 2020 Jul 6. pii: 10.1038/s41587-020-0571-7. doi: 
            10.1038/s41587-020-0571-7."
COMMENT     Sequence Label: pJBL703
FEATURES             Location/Qualifiers
     source          1..2921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..76
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    193..214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(229..234,235..252,253..259)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    267..283
                     /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)."
     primer_bind     272..294
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     291..307
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     291..307
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        350..368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    369..387
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      540..586
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(612..629)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(612..628)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(621..643)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(837..856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     956..978
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1016..1034)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1102..1206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1207..1275,1276..2067)
                     /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     complement(1425..1444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2238..2826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2727..2746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
       61 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
      121 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
      181 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
      241 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
      301 ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagat aatacgactc
      361 actataggtc cctatcagtg atagagaccc acatactctg atgatccgag acggtcgggt
      421 ccagatattc gtatctgtcg agtagagtgt gggctcggat cattcatggc aagagacggt
      481 cgggtccaga tattcgtatc tgtcgagtag agtgtgggct cttgccatgt gtatgtgggt
      541 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgatcc ccgggtaccg
      601 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      661 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      721 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
      781 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
      841 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
      901 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
      961 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     1021 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1081 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1141 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1201 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1261 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1321 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1381 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1441 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1501 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1561 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1621 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1681 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1741 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1801 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1861 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1921 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1981 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2041 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2101 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2161 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2221 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2281 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2341 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2521 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2761 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2821 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2881 cacatgttct ttcctgcgtt atcccctgat tctgtggata a
//
