LOCUS       sequence-102827-        5098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acrylate biosensor. Low copy plasmid that expresses GFP proportional
            to acrylate concentration..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5098)
  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 5098)
  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-L-acuR
FEATURES             Location/Qualifiers
     source          1..5098
                     /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"
     CDS             complement(1072..2022)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2070..2292
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(2784..2801)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2954..3040
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3132..3159
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             3278..3991
                     /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             4284..4292
                     /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)"
     terminator      5013..5060
                     /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 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggctctcg
      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 taacgtgagt tttcgttcca ctgagcgtca gaccccgaac ctcagatcct
     1081 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     1141 cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt
     1201 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta
     1261 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     1321 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     1381 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     1441 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     1501 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     1561 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     1621 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     1681 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     1741 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     1801 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     1861 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     1921 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     1981 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     2041 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt
     2101 cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     2161 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     2221 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     2281 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     2341 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     2401 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     2461 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     2521 actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg
     2581 tcacgggctt ctcagggcgt tttatggcgg atctgctatg tggtgctatc tgactttttg
     2641 ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac
     2701 aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac tggctgtttt
     2761 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     2821 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     2881 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     2941 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3001 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     3061 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     3121 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactcgcta
     3181 gcaagtaagg ccgactgctt cacaaccgca ctttgattta atagaccata ccgtctatta
     3241 tttctggaac atttaacttt aagaaggaga tatacatatg cgtaaaggtg aagaactgtt
     3301 caccggtgtt gttccgatcc tggttgaact ggacggtgac gttaacggtc acaaattctc
     3361 tgttcgtggt gaaggtgaag gtgacgctac caacggtaaa ctgaccctga aattcatctg
     3421 caccaccggt aaactgccgg ttccgtggcc gaccctggtt accaccctga cctacggtgt
     3481 tcagtgcttc gctcgttacc cggaccacat gaaacagcac gacttcttca aatctgctat
     3541 gccggaaggt tacgttcagg aacgtaccat ctctttcaaa gacgacggta cctacaaaac
     3601 ccgtgctgaa gttaaattcg aaggtgacac cctggttaac cgtatcgaac tgaaaggtat
     3661 cgacttcaaa gaagacggta acatcctggg tcacaaactg gaatacaact tcaactctca
     3721 caacgtttac atcaccgctg acaaacagaa aaacggtatc aaagctaact tcaaaatccg
     3781 tcacaacgtt gaagacggtt ctgttcagct ggctgaccac taccagcaga acaccccgat
     3841 cggtgacggt ccggttctgt tgccggacaa ccactacctg tctacccagt ctgttctgtc
     3901 taaagacccg aacgaaaaac gtgaccacat ggttctgctg gaattcgtta ccgctgctgg
     3961 tatcacccac ggtatggacg aactgtacaa agatgcatgc cagttctagc ataaccctaa
     4021 tgagtgagct aacttacatt aattgcgttg cgccttaatt aacggcactc ctcagcaaat
     4081 ataatgaccc tcttgataac ccaagagggc attttttaat gcccatggcg tttaccacag
     4141 ctaacaccac gtcgtcccta tctgctgccc taggtctatg agtggttgct ggataacttt
     4201 acgggcatgc ataaggctcg taggctatat tcagggagac cacaacggtt tccctctaca
     4261 aataattttg tttaactttg aaataaggag gtaatacaaa tgccgctgac cgacaccccg
     4321 ccgtctgttc cgcagaaacc gcgtcgtggt cgtccgcgtg gtgctccgga cgcttctctg
     4381 gctcaccagt ctctgatccg tgctggtctg gaacacctga ccgaaaaagg ttactcttct
     4441 gttggtgttg acgaaatcct gaaagctgct cgtgttccga aaggttcttt ctaccactac
     4501 ttccgtaaca aagctgactt cggtctggct ctgatcgaag cttacgacac ctacttcgct
     4561 cgtctcctcg accaggcgtt cctggacggt tcgctggctc cgctggctcg tctgcgtctg
     4621 ttcacccgta tggctgaaga aggtatggct cgtcacggtt tccgtcgtgg ttgcctggtt
     4681 ggtaacctgg gtcaggaaat gggtgctctg ccggacgact tccgtgctgc tctgatcggt
     4741 gttctggaaa cctggcagcg tcgtaccgct cagctgttcc gtgaagctca ggcttgcggt
     4801 gaactgtctg ctgaccacga cccggacgct ctggctgaag ctttctggat cggttgggaa
     4861 ggtgctatcc tgcgtgctaa actggaactg cgtccggacc cgctgcactc tttcacccgt
     4921 accttcggtc gtcacttcgt tacccgtacc catgaataat aagcgactaa aaaattgaat
     4981 gtaggaaacc aacatgccag ttcgagcaat aactagcata accccttggg gcctctaaac
     5041 gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacact
//
