LOCUS       sequence-325107-        3901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for: roGFP2-Orp1 H2O2 oxidation sensor (cytosolic).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3901)
  AUTHORS   Werley CA, Boccardo S, Rigamonti A, Hansson EM, Cohen AE
  TITLE     Multiplexed Optical Sensors in Arrayed Islands of Cells for 
            multimodal recordings of cellular physiology.
  JOURNAL   Nat Commun. 2020 Aug 4;11(1):3881. doi: 10.1038/s41467-020-17607-5.
  PUBMED    32753572
REFERENCE   2  (bases 1 to 3901)
  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: "Nat 
            Commun."; date: "2020-08-4"; volume: "11(1):3881. doi"; pages: " 
            10.1038/s41467-020-17607-5"
COMMENT     Sequence Label: pMOS016E: Entry vector for: roGFP2-Orp1 H2O2
            o...
FEATURES             Location/Qualifiers
     source          1..3901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(707..709,710..712,713..1423)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(752..773)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1013..1032)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1360..1381
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2026..2125)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2142..2161)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2143..2161)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2166..2182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2166..2182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2284..2303)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2295..3104
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2362..2381)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3251..3839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3740..3759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttt aaaggaacca attcagtcga ctggatctgc ggccgcatgg tgagcaaggg
      721 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      781 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      841 gaagttcatc tccaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      901 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      961 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1021 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1081 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1141 ctacaactgc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1201 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1261 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcacctg
     1321 ctccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1381 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagactagtg gtggttcagg
     1441 tggtggtggt tcaggtggtg gtggttcagg tggaggagga tcaggaggag gaggatcagg
     1501 aggaggagga tcaggaggag aattcgatat ctcagaattc tataagctag cacctgttga
     1561 caagaaaggc caaccattcc ccttcgacca attaaaggga aaagtggtgc ttatcgttaa
     1621 tgttgcctcc aaatgtggat tcactcctca atacaaagaa ctagaggcct tgtacaaacg
     1681 ttataaggac gaaggattta ccatcatcgg gttcccatgc aaccagtttg gccaccaaga
     1741 acctggctct gatgaagaaa ttgcccagtt ctgccaactg aactatggcg tgactttccc
     1801 cattatgaaa aaaattgacg ttaatggtgg caatgaggac cctgtttaca agtttttgaa
     1861 gagccaaaaa tccggtatgt tgggcttgag aggtatcaaa tggaattttg aaaaattctt
     1921 agtcgataaa aagggtaaag tgtacgaaag atactcttca ctaaccaaac cttcttcgtt
     1981 gtccgaaacc atcgaagaac ttttgaaaga ggtggaatag tctagaccca gctttcttgt
     2041 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     2101 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     2161 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     2221 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     2281 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     2341 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2401 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2461 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2521 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2581 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2641 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     2701 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2761 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2821 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2881 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2941 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     3001 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     3061 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     3121 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     3181 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     3241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3361 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3841 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3901 t
//
