LOCUS       sequence-325091-        3947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for: Mitycam calcium sensor (mitochondrial).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3947)
  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 3947)
  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: pMOS028E: Entry vector for: Mitycam calcium se...
FEATURES             Location/Qualifiers
     source          1..3947
                     /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             672..746
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             783..860
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             903..1154
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSFQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1091..1112
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1224..1245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1485..1504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(2072..2171)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2188..2207)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2189..2207)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2212..2228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2212..2228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2330..2349)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2341..3150
                     /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(2408..2427)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3297..3885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3786..3805
                     /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 agcaggctac catgtccgtc ctgacgccgc tgctgctgcg gggcttgaca ggctcggccc
      721 ggcggctccc agtgccgcgc gccaagatcc attcgttgcc gccggagggg aagcttggca
      781 tgaagaggcg ctggaagaaa aacttcattg ccgtcagcgc tgccaaccgg ttcaagaaga
      841 tctccagctc cggggcactg gggtctgcag gctacaacag caccaacgtc tatatcatgg
      901 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
      961 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1021 tgctgcccga caaccactac ctgagcttcc agtccgccct gagcaaagac cccaacgaga
     1081 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1141 acgagctgta caaggtcgac ggtggcagcg gtggcaccgg tgtgagcaag ggcgaggagc
     1201 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1261 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctca
     1321 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
     1381 gcctgaagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1441 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1501 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1561 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacggtaccg
     1621 accaactgac agaagagcag attgcagagt tcaaagaagc cttctcatta ttcgacaagg
     1681 atggggacgg caccatcacc acaaaggaac ttggcaccgt tatgaggtcg cttggacaaa
     1741 acccaacgga agcagaattg caggatatga tcaatgaagt cgatgctgat ggcaatggaa
     1801 cgatttactt tcctcaattt cttactatga tggctagaaa aatgaaggac acagacagcg
     1861 aagaggaaat ccgagaagca ttccgtgttt ttgacaagga tgggaacggc tacatcagcg
     1921 ctgctgaatt acgtcacgtc atgacaaacc tcggggagaa gttaacagat gaagaagttg
     1981 atgaaatgat aagggaagca gatatcgatg gtgatggcca agtaaactat gaagagtttg
     2041 tacaaatgat gacagcaaag taagcggccg cacccagctt tcttgtacaa agttggcatt
     2101 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     2161 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     2221 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2281 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2341 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2401 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2461 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2521 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2581 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2641 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2701 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2761 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2821 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2881 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2941 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     3001 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     3061 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     3121 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     3181 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3241 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3301 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3361 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3421 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3481 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3541 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3601 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3661 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3721 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3781 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3841 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3901 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
