LOCUS       sequence-409924-        4249 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression of the red fluorescent rogRFP2 redox sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4249)
  AUTHORS   Radhakrishnan S, Norley J, Wendt S, LeRoy N, Hall H, Norcross S, 
            Doan S, Snaider J, MacVicar BA, Weake VM, Huang L, Tantama M
  TITLE     Neuron Activity Dependent Redox Compartmentation Revealed with a 
            Second Generation Red-Shifted Ratiometric Sensor.
  JOURNAL   ACS Chem Neurosci. 2020 Sep 2;11(17):2666-2678. doi: 
            10.1021/acschemneuro.0c00342. Epub 2020 Aug 21.
  PUBMED    32786310
REFERENCE   2  (bases 1 to 4249)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acschemneuro.0c00342"; journalName: "ACS Chem Neurosci"; 
            date: "2020-09-2- 2"; volume: "11"; issue: "17"; pages: "2666-2678"
COMMENT     Sequence Label: pRset-rogRFP2
FEATURES             Location/Qualifiers
     source          1..4249
                     /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"
     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
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(247..268)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(508..527)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     855..876
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1290..1309
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1650..1668)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1664..1711
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1808..2263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1895..1914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2105..2126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2289..2393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2394..2462,2463..3254)
                     /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(2612..2631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3425..4013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3914..3933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4167..4184
                     /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 aggatcccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      241 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      301 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctccac caccggcaag
      361 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca 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 actgccacaa cgtctatatc
      661 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
      721 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      781 gtgctgctgc ccgacaacca ctacctgagc acctgctccg ccctgagcaa agaccccaac
      841 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ctgcaggtgg agaggaaaac
      901 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
      961 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggc ctttcagacc
     1021 gctaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     1081 cagttcatgt acggctccaa ggtctacatt aagcacccag ccgacatccc cgactacttc
     1141 aagctgtcct tccccgaggg cttcaggtgg gagcgcgtga tgaacttcga ggacggcggc
     1201 attattcacg ttaaccagga ctcctccctg caggacggcg tgttcatcta caaggtgaag
     1261 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     1321 gaggcctccg aggagcggat gtaccccgag gacggcgccc tgaagagcga gatcaagaag
     1381 aggctgaagc tgaaggacgg cggccactac gccgccgagg tcaagaccac ctacaaggcc
     1441 aagaagcccg tgcagctgcc cggcgcctac atcgtcgaca tcaagttgga catcgtgtcc
     1501 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggtcg tcactctacc
     1561 ggcggcatgg acgagctgta caagtaagaa ttcgaagctt gatccggctg ctaacaaagc
     1621 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1681 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatctgg
     1741 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1801 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1861 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1921 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1981 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2041 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2101 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2161 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2221 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     2281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3541 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4201 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcag
//
