LOCUS       sequence-251585-        4644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Unmodified FLIP CitA6-130 sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4644)
  AUTHORS   Ewald JC, Reich S, Baumann S, Frommer WB, Zamboni N
  TITLE     Engineering genetically encoded nanosensors for real-time in vivo 
            measurements of citrate concentrations.
  JOURNAL   PLoS One. 2011;6(12):e28245. doi: 10.1371/journal.pone.0028245. Epub
            2011 Dec 2.
  PUBMED    22164251
REFERENCE   2  (bases 1 to 4644)
  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: "PLoS One.";
            date: "2011"; volume: "6(12):e28245. doi"; pages: " 
            10.1371/journal.pone.0028245. Epub 2011 Dec 2"
COMMENT     Sequence Label: CFP-CitA6-130-Venus
FEATURES             Location/Qualifiers
     source          1..4644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     200..219
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        200..218
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             250..272
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             292..309
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             313..345
                     /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(349..357,358..372)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(388..390,391..903)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIE"
     CDS             1915..2166
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     complement(2225..2243)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2239..2286
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2383..2838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2470..2489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2680..2701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2864..2968
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2969..3037,3038..3829)
                     /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(3187..3206)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4000..4588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4489..4508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      241 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      301 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      361 gatgacgata aggatccggg cctcgaggtg agcaagggcg aggagctgtt caccggggtg
      421 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      481 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      541 aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc
      601 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      661 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      721 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      781 gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat
      841 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
      901 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
      961 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1021 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gggtacccgt
     1081 ctgcattatc aggtcgggca gcgggcgctg attcaggcaa tgcagatttc ggcgatgccg
     1141 gagctggtcg aagcggttca gaaacgcgat ctcgccagaa tcaaagcgct tatcgatccc
     1201 atgcgttcgt tctccgacgc cacctatatc accgtcggtg atgccaacgg ccagcgcctc
     1261 tatcatgtta atcctgatga aatcggcaaa tcgatggagg gcggcgatag cgatgaagca
     1321 ttgttaaacg cgaaaagtta cgtgtcagtg cgcaaaggct cgctgggctc ctcgctgcgc
     1381 ggtaaatcac cgattcagga cgagaccggc aaggtgattg gcattgtgtc ggtaggctat
     1441 accatcgagc aaactagtaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1501 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1561 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     1621 tggcccaccc tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac
     1681 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1741 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1801 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1861 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     1921 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     1981 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2041 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2101 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2161 tacaagtaag ggcccgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     2221 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     2281 tttttgctga aaggaggaac tatatccgga tctggcgtaa tagcgaagag gcccgcaccg
     2341 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     2401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2581 accccaaaaa acttgattag ggtgttggtt cacgtagtgg gccatcgccc tgatagacgg
     2641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2761 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     2821 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2881 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2941 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3001 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3061 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3121 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3181 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3241 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3301 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3361 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3421 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3481 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3541 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3601 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3661 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3721 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3781 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3841 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3901 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3961 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4021 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4081 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4141 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4201 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4261 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4321 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4381 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4441 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4501 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4561 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4621 ggccttttgc tggccttttg ctca
//
