LOCUS       sequence-390928-        5387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes dual cistronic nuclear localized GCaMP6s and mScarlet-I 
            optimized for C. elegans expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5387)
  TITLE     plasmids optimized for C. elegans expression
REFERENCE   2  (bases 1 to 5387)
  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"
COMMENT     Sequence Label: pDSP23
FEATURES             Location/Qualifiers
     source          1..5387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     53..69
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             227..247
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             263..280
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(320..328,329..343)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             371..427
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             2122..2142
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      3087..3410
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(3508..3524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3508..3524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(3615..4203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3695..3714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4374..5189)
                     /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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5097..5116
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5190..5292)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     5360..5378
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagtaaaacg
       61 acggccagtt tatctagtca gcttgattct agctgatcgt ggaccggaag gtgagccagt
      121 gagttgattg cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatgcggccg
      181 catggtttcc aagggagagg ctgttatcaa ggaattcatg acttacccaa agaagaagcg
      241 taaggtccag gttatgggat cccaccacca ccaccaccac ggaatggcct ctatgaccgg
      301 aggacaacaa atgggacgcg acctttacga tgacgacgac aaggaccttg ctaccatggt
      361 tgattcttct cgccgcaagt ggaacaagac tggacatgct gtccgtgcca tcggacgtct
      421 ttcctccctt gagaacgttt acatcaaggc tgataagcaa aagaacggaa ttaaggctaa
      481 cttccatatc cgtcacaaca tcgaagatgg aggagtccaa cttgcttacc attaccaaca
      541 aaacactcca atcggagatg gaccagttct tcttccagat aaccactacc tttccgttca
      601 atccaagctc tctaaggtaa gtttaaacat atatatacta actaaccctg attatttaaa
      661 ttttcaggat ccaaacgaga agcgtgacca catggtcctt cttgagttcg tcaccgctgc
      721 tggaatcacc cttggaatgg atgagcttta taagggagga actggaggat ctatggtttc
      781 taagggagag gagcttttca ctggagttgt cccaatcctt gtcgagcttg atggagatgt
      841 taacggacac aagttctctg tttctggaga gggagaggga gacgctacct acggaaagct
      901 tactcttaag ttcatctgca ctactggaaa gcttccagtt ccatggccaa cccttgttac
      961 cacccttact tacggagttc aatgcttctc tcgttaccca gatcatatga agcaacatga
     1021 tttcttcaag tctgctatgc cagagggata catccaagag cgtactatct tcttcaaggt
     1081 aagtttaaac agttcggtac taactaacca tacatattta aattttcagg atgatggaaa
     1141 ctacaagacc cgtgctgagg ttaagttcga gggagatacc cttgttaacc gtatcgagct
     1201 taagggaatc gatttcaagg aagatggaaa catccttgga cataagcttg agtacaacct
     1261 tccagatcaa cttaccgagg aacaaatcgc tgagttcaag gaggccttct cccttttcga
     1321 taaggatgga gacggaacta tcactaccaa ggtaagtttt ctagagacct ttactaacta
     1381 actaggtctc atagattttc aggagcttgg aaccgttatg cgttcccttg gacaaaaccc
     1441 aactgaggct gagcttcaag atatgatcaa cgaggttgac gctgatggag atggaactat
     1501 cgatttccca gagttcctta ctatgatggc tcgcaagatg aagtaccgcg atactgagga
     1561 ggagatccgc gaggctttcg gagtcttcga caaggacgga aacggataca tttccgctgc
     1621 tgagctccgt cacgttatga ctaacctcgg agagaagctt accgacgagg aggtcgacga
     1681 gatgatccgt gaggctgata tcgacggaga cggacaagtt aactacgagg agttcgttca
     1741 aatgatgact gccaagtccc gtacgcgtaa ggccaaccca accaagctct ccgagaacgc
     1801 caagaagctc gccaaggagg tcgagaacta agctgtctca tcctactttc acctagttaa
     1861 ctgcttgtct taaaatctat gcttctcttt agtatctaaa attttcctag aagcttacaa
     1921 gtatataaat ggtctcttct caataaaggt tgtatattta ttcatcttat tgaatctgcc
     1981 atttcctcgt ttttgcgagt ttatatacct tccaattttc tttctattgt attttcaact
     2041 tctaatttta attcagggaa actgcttcaa cgcatcatgg tttccaaggg agaggctgtt
     2101 atcaaggaat tcatgactta cccaaagaag aagcgtaagg tcgaacttat caaggagttt
     2161 atgcgcttta aggttcacat ggaaggatct atgaacggac acgaattcga aatcgaagga
     2221 gaaggagaag gacgtccata cgagggaact caaactgcta agcttaaggt aagtttatct
     2281 aaaaagtttt tcattcaaaa tgtgtaaaaa ttcatttaaa ataacccaaa aatcattaat
     2341 cctcgatatt ttcaggttac taaaggagga ccacttccat tctcttggga tatcctttct
     2401 ccacaattca tgtacggatc tcgcgctttc atcaagcacc cagctgatat cccagattac
     2461 tacaagcaat ctttcccaga aggattcaaa tgggagcgtg ttatgaactt cgaagatgga
     2521 ggagctgtta ccgttaccca agatacctcc cttgaggatg gaacccttat ctacaaggta
     2581 ttttcctgca tttttcaact gggaaaatga aagaaaatcg ataatttcag gttaagcttc
     2641 gtggaactaa tttcccacca gatggaccag ttatgcaaaa gaagactatg ggatgggagg
     2701 cttctaccga gcgcctttac ccagaggatg gagtccttaa gggagatatc aagatggctc
     2761 ttcgtcttaa ggatggagga cgttaccttg ctgatttcaa ggtaagtttt ctagagacct
     2821 ttactaacta actaggtctc atagattttc agactactta caaggctaag aagccagttc
     2881 aaatgccggg agcttacaac gttgatcgta agcttgatat cacctctcac aacgaggatt
     2941 acaccgttgt tgagcaatac gagcgttctg agggacgtca ctctaccgga ggaatggatg
     3001 agctttacaa gtcccgtacg cgtaaggcca acccaaccaa gctctccgag aacgccaaga
     3061 agctcgccaa ggaggtcgag aactaaatgc aaaatccttt caagcattcc cttcttctct
     3121 atcactcttc tttctttttg tcaaaaaatt ctctcgctaa tttatttgct tttttaatgt
     3181 tattatttta tgacttttta tagtcactga aaagtttgca tctgagtgaa gtgaatgcta
     3241 tcaaaatgtg attctgtctg atgtactttc acaatctctc ttcaattcca ttttgaagtg
     3301 ctttaaaccc gaaaggttga gaaaaatgcg agcgctcaaa tatttgtatt gtgttcgttg
     3361 agtgacccaa caaaaagagg aaactttatt gtgccgccaa gaaaaaagtc tcaatcagtt
     3421 ctggaccagc gagctgtgct gcgatgatct tacggcatta tacgtatgat cggtccacga
     3481 tcagctagat tatctagtca gcttgatgtc atagctgttt cctgacatgt gagcaaaagg
     3541 ccagcaaaag gccaggaacc gtaaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3721 agctccctcg tgcgctctcc tgttccgacc ctgtcgctta ccggatacct gtccgccttt
     3781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttagaaa
     4381 aactcatcga gcatcaagtg aaactgcaat ttattcatat caggattatc aataccatat
     4441 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     4501 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     4561 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     4621 ggtgagaatg gcaagagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     4681 cgctcgtcat caaaatcact cgcaccaacc aaaccgttat tcattcgtga ttgcgcctga
     4741 gcgagacgaa atacgcgatc gccgttaaaa ggacaattac aaacaggaat cgaatgcaac
     4801 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     4861 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
     4921 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagcctg
     4981 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     5041 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     5101 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcttcaag
     5161 caagacgttt cccgttgaat atggctcatt ttagcttcct tagctcctga aaatctcgat
     5221 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     5281 acgtgccgat cacacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5341 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
