LOCUS       sequence-7685-ex        5218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5218)
  AUTHORS   Deuschle K, Okumoto S, Fehr M, Looger LL, Kozhukh L, Frommer WB
  TITLE     Construction and optimization of a family of genetically encoded 
            metabolite sensors by semirational protein engineering.
  JOURNAL   Protein Sci. 2005 Sep . 14(9):2304-14.
  PUBMED    16131659
REFERENCE   2  (bases 1 to 5218)
  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: "Protein 
            Sci. 2005 Sep . 14(9):2304-14."
COMMENT     Sequence Label: pRSET FLIPglu-3.2mDelta13
FEATURES             Location/Qualifiers
     source          1..5218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             67..89
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             109..126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             130..162
                     /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(166..174,175..189)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(202..204,205..207,208..720)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             2254..2505
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     complement(2564..2582)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2578..2625
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2722..3177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2809..2828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3019..3040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3258..3362
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3363..3431,3432..4223)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4394..4982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4883..4902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5136..5153
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gataattttg tttaacttta agaaggagat atacatatgc ggggttctca tcatcatcat
      121 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      181 gacgataagg atccgggccg catggtgagc aagggcgagg agctgttcac cggggtggtg
      241 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      301 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      361 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
      421 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      481 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      541 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      601 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
      661 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      721 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      781 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
      841 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cgctgatact
      901 cgcattggtg taacaatcta taagtacgac gataacgcga tgtctgtagt gcgcaaggct
      961 attgagcaag atgcgaaagc cgcgccagat gttcagctgc tgatgaatga ttctcagaat
     1021 gaccagtcca agcagaacga tcagatcgac gtattgctgg cgaaaggggt gaaggcactg
     1081 gcaatcaacc tggttgaccc ggcagctgcg ggtacggtga ttgagaaagc gcgtgggcaa
     1141 aacgtgccgg tggttttctt caacaaagaa ccgtctcgta aggcgctgga tagctacgac
     1201 aaagcctact acgttggcac tgacgccaaa gagtccggca ttattcaagg cgatttgatt
     1261 gctaaacact gggcggcgaa tcagggttgg gatctgaaca aagacggtca gattcagttc
     1321 gtactgctga aaggtgaacc gggccatccg gatgcagaag cacgtaccac ttacgtgatt
     1381 aaagaattga acgataaagg catcaaaact gaacagttac agttagatac cgcaatgtgg
     1441 gacaccgctc aggcgaaaga taagatggac gcctggctgt ctggcccgaa cgccaacaaa
     1501 atcgaagtgg ttatcgccaa caacgatgcg atggcaatgg gcgcggttga agcgctgaaa
     1561 gcacacaaca agtccagcat tccggtgttt ggcgtcgatg cgctgccaga agcgctggcg
     1621 ctggtgaaat ccggtgcact ggcgggcacc gtactgaacg atgctaacaa ccaggcgaaa
     1681 gcgacctttg atctggcgaa aaacctggcc gatggtaaag gtgcggctga tggcaccaac
     1741 tggaaaatcg acaacaaagt ggtccgcgta ccttatgttg gcgtagataa agacaacctg
     1801 gctctcgagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1861 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1921 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1981 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2041 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2101 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2161 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2221 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2281 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2341 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     2401 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2461 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaaa gcttgatccg
     2521 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     2581 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     2641 atatccggat ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2701 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2761 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2821 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2881 tccctttagg gttccgattt agagctttac ggcacctcga ccgcaaaaaa cttgatttgg
     2941 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3001 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatcg
     3061 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3121 agctgattta acaaatattt aacgcgaatt ttaacaaaat attaacgttt acaatttcgc
     3181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acaggtggca
     3241 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3301 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3361 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3421 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3481 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3541 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtgat acactattat
     3601 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3661 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3721 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3781 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3841 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagagt gacaccacga
     3901 tgcctgtagc aatgccaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3961 cttcccggca acaattaata gactgaatgg aggcggataa agttgcagga ccacttctgc
     4021 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4081 ctcgcggtat cattgcagca ctggggccag atggtaagcg ctcccgtatc gtagttatct
     4141 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4201 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4261 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4321 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4381 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4441 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4501 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     4561 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4621 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4681 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4741 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4801 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4861 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4921 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4981 aaaacgccag caacgcggcc tttttacggt tcctgggctt ttgctggcct tttgctcaca
     5041 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5101 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5161 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcag
//
