LOCUS       sequence-39933-e        4750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a single-wavelength maltose sensor constructed from E. coli MalE and
            cpGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4750)
  AUTHORS   Marvin JS, Schreiter ER, Echevarria IM, Looger LL
  TITLE     A genetically encoded, high-signal-to-noise maltose sensor.
  JOURNAL   Proteins. 2011 Nov;79(11):3025-36. doi: 10.1002/prot.23118.
  PUBMED    21989929
REFERENCE   2  (bases 1 to 4750)
  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"; doi: "10.1002/prot"; 
            journalName: "Proteins"; date: "2011-11"; volume: "79"; issue: "11";
            pages: "3025-36"
COMMENT     Sequence Label: EcMBP165-cpGFP.PPYF.T203V.pRSET
FEATURES             Location/Qualifiers
     source          1..4750
                     /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             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     918..939
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1045..1066)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1306..1325)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2065..2082
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2151..2169)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2165..2212
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2309..2764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2396..2415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2606..2627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2790..2894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2895..2963,2964..3755)
                     /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(3113..3132)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3926..4514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4415..4434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4668..4685
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatcgatg gggatccaaa atcgaagaag gtaaactggt aatctggatt
      241 aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc
      301 ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc acaggttgcg
      361 gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct
      421 caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg
      481 tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa
      541 gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac ctgggaagag
      601 atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg
      661 caagaaccgt acttcacctg gccgctgatt gctgctgacc cccccagcta caacgtcttc
      721 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
      781 gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat cggcgacggc
      841 cccgtgctgc tgcccgacaa ccactacctg agcgtccagt ccaaactttc gaaagacccc
      901 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
      961 ggcatggacg agctgtacaa gggcggtacc ggagggagca tggtgagcaa gggcgaggag
     1021 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1081 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1141 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1201 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1261 gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac
     1321 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1381 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactttaac
     1441 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
     1501 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
     1561 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
     1621 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     1681 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     1741 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     1801 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     1861 aagtcttacg aggaagagtt ggtgaaagat ccgcggattg ccgccactat ggaaaacgcc
     1921 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     1981 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagacct gaaagacgcg
     2041 cagactcgga tcaccaaagg aagtcaccat catcaccacc atggctaaga attcgaagct
     2101 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2161 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     2221 aggaactata tccggatctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2281 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     2341 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2401 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2461 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2521 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2581 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2641 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2701 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     2761 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2821 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2881 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2941 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3001 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3061 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3121 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3181 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3241 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3301 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3361 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3421 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3481 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3541 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3601 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3661 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3721 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3781 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3841 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3901 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3961 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4021 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4081 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4141 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4201 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4261 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4321 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4381 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4441 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4501 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4561 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4621 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4681 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4741 attaatgcag
//
