LOCUS       sequence-315859-        4164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for: QuasAr2-mOrange2 voltage sensor (plasma membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4164)
  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 4164)
  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: pMOS009E: Entry vector for: QuasAr2-mOrange2 v...
FEATURES             Location/Qualifiers
     source          1..4164
                     /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             703..1461
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     CDS             1468..2175
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEYNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     protein_bind    complement(2289..2388)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2405..2424)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2406..2424)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2429..2445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2429..2445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2547..2566)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2558..3367
                     /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(2625..2644)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3514..4102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4003..4022
                     /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 agcaggctgg cgccggaacc aattcagtcg actggatcca ccatggtaag tatcgctctg
      721 caggctggtt acgacctact gggtgacggc agacctgaaa ccctgtggct gggcatcggc
      781 actctgctga tgctgattgg aaccttctac tttctggtcc gcggatgggg agtcaccgat
      841 aaggatgccc gggaatatta cgctgtgact atcctggtgt ccggaatcgc atccgccgca
      901 tatctgtcta tgttctttgg tatcgggctt actgaggtgt ccgtcggggg cgaaatgttg
      961 gatatctatt atgccaggta cgcccagtgg ctgtttacca ccccacttct gctgctgcac
     1021 ctggcccttc tcgctaaggt ggatcgggtg accatcggca ccctggtggg tgtggacgcc
     1081 ctgatgatcg tcactggcct catcggagcc ttgagccaca cggccatagc cagatacagt
     1141 tggtggttgt tctctacaat ttgcatgata gtggtgctct atgttctggc tacatccctg
     1201 cgatctgctg caaaggagcg gggccccgag gtggcatcta cctttaacac cctgacagct
     1261 ctggtcttgg tgctgtggac cgcttaccct atcctgtgga tcataggcac tgagggcgct
     1321 ggcgtggtgg gcctgggcat cgaaactctg ctgtttatgg tgttggacgt gactgccaag
     1381 gtcggctttg gctttatcct gttgagatcc cgggctattc tgggcgacac cgaggcacca
     1441 gaacccagtg ccggtgccga tacgttgatg gtgagcaagg gcgaggagta taacatggcc
     1501 atcatcaagg agttcatgcg cttcaaggtg cgcatggagg gctccgtgaa cggccacgag
     1561 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gctttcagac cgctaagctg
     1621 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcatttcacc
     1681 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt caagctgtcc
     1741 ttccccgagg gcttcaagtg ggagcgcgtg atgaactacg aggacggcgg cgtggtgacc
     1801 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1861 accaacttcc cctccgacgg ccccgtgatg cagaagaaga ccatgggctg ggaggcctcc
     1921 tccgagcgga tgtaccccga ggacggtgcc ctgaagggca agatcaagat gaggctgaag
     1981 ctgaaggacg gcggccacta cacctccgag gtcaagacca cctacaaggc caagaagccc
     2041 gtgcagctgc ccggcgccta catcgtcgac atcaagttgg acatcacctc ccacaacgag
     2101 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2161 gacgagctgt acaagagtag aatcacaagc gaaggcgagt acatccccct ggatcaaata
     2221 gacataaatg taggtggatt ttgttatgag aatgaataag aattcgcggc cgcactcgag
     2281 atatctagac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2341 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2401 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2461 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2521 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2581 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2641 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2701 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2761 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2821 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2881 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2941 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3001 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3061 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3121 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3181 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3241 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3301 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3361 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3421 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3661 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4141 ttgctggcct tttgctcaca tgtt
//
