LOCUS       sequence-115680-        4076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tetracycline biosensor. High copy plasmid that expresses GFP 
            proportional to tetracycline concentration..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4076)
  AUTHORS   Rogers JK, Guzman CD, Taylor ND, Raman S, Anderson K, Church GM
  TITLE     Synthetic biosensors for precise gene control and real-time 
            monitoring of metabolites.
  JOURNAL   Nucleic Acids Res. 2015 Jul 7. pii: gkv616.
  PUBMED    26152303
REFERENCE   2  (bases 1 to 4076)
  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: "Nucleic 
            Acids Res. 2015 Jul 7. pii: gkv616."
COMMENT     Sequence Label: pJKR-H-tetR
FEATURES             Location/Qualifiers
     source          1..4076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(59..127,128..919)
                     /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(277..296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1090..1678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1579..1598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1767..1784)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1937..2023
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(2185..2203,2204..2209,2210..2226,2227..2232,
                     2233..2258)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    2185..2203
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2210..2228
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2297..3010
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             3303..3311
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             3319..3942
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(3392..3411)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3996..4043
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
       61 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      661 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      901 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1021 caaaatccct taacgtgggt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1081 aggatcatct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1201 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcaggctgt
     1741 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1801 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1861 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1921 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1981 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     2041 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     2101 gcatcaaatt aagcagaagg ccatccgtca ggatggcctt tttgcgtttc tacaaactct
     2161 gctagcaagt aaggccgact cgagtcccta tcagtgatag agattgacat ccctatcagt
     2221 gatagagata ctgagcacat cagcaggacg cactgaccga attcattaaa tttaacttta
     2281 agaaggagat atacatatgc gtaaaggtga agaactgttc accggtgttg ttccgatcct
     2341 ggttgaactg gacggtgacg ttaacggtca caaattctct gttcgtggtg aaggtgaagg
     2401 tgacgctacc aacggtaaac tgaccctgaa attcatctgc accaccggta aactgccggt
     2461 tccgtggccg accctggtta ccaccctgac ctacggtgtt cagtgcttcg ctcgttaccc
     2521 ggaccacatg aaacagcacg acttcttcaa atctgctatg ccggaaggtt acgttcagga
     2581 acgtaccatc tctttcaaag acgacggtac ctacaaaacc cgtgctgaag ttaaattcga
     2641 aggtgacacc ctggttaacc gtatcgaact gaaaggtatc gacttcaaag aagacggtaa
     2701 catcctgggt cacaaactgg aatacaactt caactctcac aacgtttaca tcaccgctga
     2761 caaacagaaa aacggtatca aagctaactt caaaatccgt cacaacgttg aagacggttc
     2821 tgttcagctg gctgaccact accagcagaa caccccgatc ggtgacggtc cggttctgtt
     2881 gccggacaac cactacctgt ctacccagtc tgttctgtct aaagacccga acgaaaaacg
     2941 tgaccacatg gttctgctgg aattcgttac cgctgctggt atcacccacg gtatggacga
     3001 actgtacaaa gatgcatgcc agttctagca taaccctaat gagtgagcta acttacatta
     3061 attgcgttgc gccttaatta acggcactcc tcagcaaata taatgaccct cttgataacc
     3121 caagagggca ttttttaatg cccatggcgt ttaccacagc taacaccacg tcgtccctat
     3181 ctgctgccct aggtctatga gtggttgctg gataacttta cgggcatgca taaggctcgt
     3241 aggctatatt cagggagacc acaacggttt ccctctacaa ataattttgt ttaactttga
     3301 aataaggagg taatacaaat gtctcgttta gataaaagta aagtgattaa cagcgcatta
     3361 gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta
     3421 ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc
     3481 ttagccattg agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc
     3541 tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc
     3601 gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa
     3661 aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc
     3721 agcgctgtgg ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct
     3781 aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc
     3841 gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata
     3901 tgcggattag aaaaacaact taaatgtgaa agtgggtctt aataagcgac taaaaaattg
     3961 aatgtaggaa accaacatgc cagttcgagc aataactagc ataacccctt ggggcctcta
     4021 aacgggtctt gaggggtttt ttgctgaacc tcaggcgaga agcacacggt cacact
//
