LOCUS       sequence-102833-        4078 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Acrylate biosensor. High 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 4078)
  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 4078)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-acuR
FEATURES             Location/Qualifiers
     source          1..4078
                     /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"
     terminator      2115..2142
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2259..2972
                     /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             3265..3273
                     /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      3994..4041
                     /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 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 ccatcctgac ggatggcctt tttgcgtttc tacaaactcg
     2161 ctagcaagta aggccgacgc ttcacaaccg cacttgattt aatagaccat accgtctatt
     2221 atttctggaa catttaactt taagaaggag atatacatat gcgtaaaggt gaagaactgt
     2281 tcaccggtgt tgttccgatc ctggttgaac tggacggtga cgttaacggt cacaaattct
     2341 ctgttcgtgg tgaaggtgaa ggtgacgcta ccaacggtaa actgaccctg aaattcatct
     2401 gcaccaccgg taaactgccg gttccgtggc cgaccctggt taccaccctg acctacggtg
     2461 ttcagtgctt cgctcgttac ccggaccaca tgaaacagca cgacttcttc aaatctgcta
     2521 tgccggaagg ttacgttcag gaacgtacca tctctttcaa agacgacggt acctacaaaa
     2581 cccgtgctga agttaaattc gaaggtgaca ccctggttaa ccgtatcgaa ctgaaaggta
     2641 tcgacttcaa agaagacggt aacatcctgg gtcacaaact ggaatacaac ttcaactctc
     2701 acaacgttta catcaccgct gacaaacaga aaaacggtat caaagctaac ttcaaaatcc
     2761 gtcacaacgt tgaagacggt tctgttcagc tggctgacca ctaccagcag aacaccccga
     2821 tcggtgacgg tccggttctg ttgccggaca accactacct gtctacccag tctgttctgt
     2881 ctaaagaccc gaacgaaaaa cgtgaccaca tggttctgct ggaattcgtt accgctgctg
     2941 gtatcaccca cggtatggac gaactgtaca aagatgcatg ccagttctag cataacccta
     3001 atgagtgagc taacttacat taattgcgtt gcgccttaat taacggcact cctcagcaaa
     3061 tataatgacc ctcttgataa cccaagaggg cattttttaa tgcccatggc gtttaccaca
     3121 gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc tggataactt
     3181 tacgggcatg cataaggctc gtaggctata ttcagggaga ccacaacggt ttccctctac
     3241 aaataatttt gtttaacttt gaaataagga ggtaatacaa atgccgctga ccgacacccc
     3301 gccgtctgtt ccgcagaaac cgcgtcgtgg tcgtccgcgt ggtgctccgg acgcttctct
     3361 ggctcaccag tctctgatcc gtgctggtct ggaacacctg accgaaaaag gttactcttc
     3421 tgttggtgtt gacgaaatcc tgaaagctgc tcgtgttccg aaaggttctt tctaccacta
     3481 cttccgtaac aaagctgact tcggtctggc tctgatcgaa gcttacgaca cctacttcgc
     3541 tcgtctcctc gaccaggcgt tcctggacgg ttcgctggct ccgctggctc gtctgcgtct
     3601 gttcacccgt atggctgaag aaggtatggc tcgtcacggt ttccgtcgtg gttgcctggt
     3661 tggtaacctg ggtcaggaaa tgggtgctct gccggacgac ttccgtgctg ctctgatcgg
     3721 tgttctggaa acctggcagc gtcgtaccgc tcagctgttc cgtgaagctc aggcttgcgg
     3781 tgaactgtct gctgaccacg acccggacgc tctggctgaa gctttctgga tcggttggga
     3841 aggtgctatc ctgcgtgcta aactggaact gcgtccggac ccgctgcact ctttcacccg
     3901 taccttcggt cgtcacttcg ttacccgtac ccatgaataa taagcgacta aaaaattgaa
     3961 tgtaggaaac caacatgcca gttcgagcaa taactagcat aaccccttgg ggcctctaaa
     4021 cgggtcttga ggggtttttt gctgaacctc aggcatttga gaagcacacg gtcacact
//
