LOCUS       sequence-39892-e        4456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a single-wavelength phosphonate sensor constructed from E. coli PhnD
            and cpGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4456)
  AUTHORS   Alicea I, Marvin JS, Miklos AE, Ellington AD, Looger LL, Schreiter 
            ER
  TITLE     Structure of the Escherichia coli Phosphonate Binding Protein PhnD 
            and Rationally Optimized Phosphonate Biosensors.
  JOURNAL   J Mol Biol. 2011 Oct 12.
  PUBMED    22019591
REFERENCE   2  (bases 1 to 4456)
  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: "J Mol Biol.
            2011 Oct 12."
COMMENT     Sequence Label: EcPhnD90.L1∆∆AD.L297RL301R.pRSET
FEATURES             Location/Qualifiers
     source          1..4456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             103..120
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     603..624
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(730..751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(991..1010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1857..1875)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1871..1918
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2015..2470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2102..2121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2312..2333
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2496..2600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2601..2669,2670..3461)
                     /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(2819..2838)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3632..4220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4121..4140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4374..4391
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcatcatca ccaccaccat
      121 ggatccgaag aacaagaaaa agctctgaac ttcggtatca tctccactga aagccaacag
      181 aacctgaagc cgcagtggac cccgttcctg caggatatgg agaagaaact gggcgttaaa
      241 gtcaacgctt tcttcgctcc ggactacgca ggcatcatcc agggtatgcg tttcaacaaa
      301 gtcgatatcg cctggtacgg caatctgtct gcaatggaag cggtcgatcg tgccaacggt
      361 caggtgttcg cgcagacggt tgcagcggac gcggacaacg tctatatcat ggccgacaag
      421 cagaggaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg
      481 cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
      541 gacaaccact acctgagcac ccagtccaaa ctttcgaaag accccaacga gaagcgcgat
      601 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
      661 tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg
      721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      841 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
      901 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      961 tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1081 gaggacggca acatcctggg gcacaagctg gagtacaact ttaacccggg ctattggtcc
     1141 gtgctgatcg ttaacaaaga cagcccgatt aacaatctga acgacctgct ggcaaaacgc
     1201 aaggacctga ccttcggcaa cggtgatcct aactccacgt ctggcttcct ggtgccaggt
     1261 tactacgtct tcgccaaaaa caacatctcc gcctctgact tcaaacgtac cgtaaacgca
     1321 ggtcacgaaa ccaacgctct ggctgttgct aacaagcagg tggacgtggc gactaataac
     1381 acggaaaacc tggacaagct gaaaaccagc gctccggaaa aactgaaaga actgaaagta
     1441 atctggaaaa gcccgctgat cccgggtgac cctatcgtat ggcgtaaaaa cctgtccgaa
     1501 accactaaag acaaaattta tgactttttc atgaactacg gtaaaacccc ggaagaaaag
     1561 gctgtgctgg aacgtctggg ctgggctccg ttccgtgctt ccagcgacct gcagctggtt
     1621 ccaatccgcc agctggctct gttcaaagaa atgcagtctg ttaaagataa caagggtctg
     1681 aacgaacagg acaagctggc taagactact gcgatccagg cacagctgga cgatctggat
     1741 cgtaggaata atgcaaggag cgcaatgtct tccgttagca aagcggtcca gtaagaattc
     1801 gaagcttgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1861 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1921 gaaaggagga actatatccg gatctggcgt aatagcgaag aggcccgcac cgatcgccct
     1981 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     2041 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2101 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2161 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2221 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2281 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2341 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2401 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2461 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2521 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2581 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2641 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2701 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2761 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2821 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2881 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2941 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3001 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3061 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3121 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3181 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3241 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3301 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3361 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3421 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3481 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3541 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3601 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3661 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3721 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3781 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3841 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3901 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3961 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4021 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4081 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4141 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4201 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4261 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4321 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4381 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4441 cgattcatta atgcag
//
