LOCUS       sequence-391045-        5169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes dual cistronic membrane localized GCaMP6f and mScarlet-I 
            optimized for C. elegans expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  TITLE     plasmids optimized for C. elegans expression
REFERENCE   2  (bases 1 to 5169)
  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: pSiM2
FEATURES             Location/Qualifiers
     source          1..5169
                     /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             191..208
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(248..256,257..271)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             299..355
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     terminator      2916..3239
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(3290..3306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3290..3306)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(3397..3985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3477..3496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4156..4971)
                     /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     4879..4898
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4972..5074)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     5142..5160
                     /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 catgggatcc caccaccacc accaccacgg aatggcctct atgaccggag gacaacaaat
      241 gggacgcgac ctttacgatg acgacgacaa ggaccttgct accatggttg attcttctcg
      301 ccgcaagtgg aacaagactg gacatgctgt ccgtgccatc ggacgtcttt cctcccttga
      361 gaacgtttac atcaaggctg ataagcaaaa gaacggaatt aaggctaact tcaagatccg
      421 tcacaacatc gaagatggag gagtccaact tgcttaccat taccaacaaa acactccaat
      481 cggagatgga ccagttcttc ttccagataa ccactacctt tccgttcaat ccaagctctc
      541 taaggtaagt ttaaacatat atatactaac taaccctgat tatttaaatt ttcaggatcc
      601 aaacgagaag cgtgaccaca tggtccttct tgagttcgtc accgctgctg gaatcaccct
      661 tggaatggat gagctttata agggaggaac tggaggatct atggtttcta agggagagga
      721 gcttttcact ggagttgtcc caatccttgt cgagcttgat ggagatgtta acggacacaa
      781 gttctctgtt tctggagagg gagagggaga cgctacctac ggaaagctta ctcttaagtt
      841 catctgcact actggaaagc ttccagttcc atggccaacc cttgttacca cccttactta
      901 cggagttcaa tgcttctctc gttacccaga tcatatgaag caacatgatt tcttcaagtc
      961 tgctatgcca gagggataca tccaagagcg tactatcttc ttcaaggtaa gtttaaacag
     1021 ttcggtacta actaaccata catatttaaa ttttcaggat gatggaaact acaagacccg
     1081 tgctgaggtt aagttcgagg gagataccct tgttaaccgt atcgagctta agggaatcga
     1141 tttcaaggaa gatggaaaca tccttggaca taagcttgag tacaaccttc cagatcaact
     1201 taccgaggaa caaatcgctg agttcaagga ggaattctcc cttttcgata aggatggaga
     1261 cggaactatc actaccaagg taagttttct agagaccttt actaactaac taggtctcat
     1321 agattttcag gagcttggaa ccgttatgcg ttcccttgga caaaacccaa ctgaggctga
     1381 gcttcaagat atgatcaacg aggttgacgc tgatggagat ggaactatcg atttcccaga
     1441 gttccttact atgatggctc gcaagatgaa gtaccgcgat actgaggagg agatccgcga
     1501 ggctttcgga gtcttcgaca aggacggaaa cggatacatt tccgctgctg agctccgtca
     1561 cgttatgact aacctcggag agaagcttac cgacgaggag gtcgacgaga tgatccgtga
     1621 ggctgatatc gacggagacg gacaagttaa ctacgaggag ttcgttcaaa tgatgactgc
     1681 caagaaacga aaggataagg gaaaatgttt aatttcctaa gctgtctcat cctactttca
     1741 cctagttaac tgcttgtctt aaaatctatg cttctcttta gtatctaaaa ttttcctaga
     1801 agcttacaag tatataaatg gtctcttctc aataaaggtt gtatatttat tcatcttatt
     1861 gaatctgcca tttcctcgtt tttgcgagtt tatatacctt ccaattttct ttctattgta
     1921 ttttcaactt ctaattttaa ttcagggaaa ctgcttcaac gcatcatggt ttccaaggga
     1981 gaggctgtta tcaaggaatt catgacttac gaacttatca aggagtttat gcgctttaag
     2041 gttcacatgg aaggatctat gaacggacac gaattcgaaa tcgaaggaga aggagaagga
     2101 cgtccatacg agggaactca aactgctaag cttaaggtaa gtttatctaa aaagtttttc
     2161 attcaaaatg tgtaaaaatt catttaaaat aacccaaaaa tcattaatcc tcgatatttt
     2221 caggttacta aaggaggacc acttccattc tcttgggata tcctttctcc acaattcatg
     2281 tacggatctc gcgctttcat caagcaccca gctgatatcc cagattacta caagcaatct
     2341 ttcccagaag gattcaaatg ggagcgtgtt atgaacttcg aagatggagg agctgttacc
     2401 gttacccaag atacctccct tgaggatgga acccttatct acaaggtatt ttcctgcatt
     2461 tttcaactgg gaaaatgaaa gaaaatcgat aatttcaggt taagcttcgt ggaactaatt
     2521 tcccaccaga tggaccagtt atgcaaaaga agactatggg atgggaggct tctaccgagc
     2581 gcctttaccc agaggatgga gtccttaagg gagatatcaa gatggctctt cgtcttaagg
     2641 atggaggacg ttaccttgct gatttcaagg taagttttct agagaccttt actaactaac
     2701 taggtctcat agattttcag actacttaca aggctaagaa gccagttcaa atgccgggag
     2761 cttacaacgt tgatcgtaag cttgatatca cctctcacaa cgaggattac accgttgttg
     2821 agcaatacga gcgttctgag ggacgtcact ctaccggagg aatggatgag ctttacaaga
     2881 aacgaaagga taagggaaaa tgtttaattt cctaaatgca aaatcctttc aagcattccc
     2941 ttcttctcta tcactcttct ttctttttgt caaaaaattc tctcgctaat ttatttgctt
     3001 ttttaatgtt attattttat gactttttat agtcactgaa aagtttgcat ctgagtgaag
     3061 tgaatgctat caaaatgtga ttctgtctga tgtactttca caatctctct tcaattccat
     3121 tttgaagtgc tttaaacccg aaaggttgag aaaaatgcga gcgctcaaat atttgtattg
     3181 tgttcgttga gtgacccaac aaaaagagga aactttattg tgccgccaag aaaaaagtct
     3241 catacgtatg atcggtccac gatcagctag attatctagt cagcttgatg tcatagctgt
     3301 ttcctgacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccagcaaaa
     3361 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3421 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3481 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgtcgct
     3541 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3601 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3661 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3721 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3781 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3841 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3901 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3961 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4021 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4081 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4141 aacttggtct gacagttaga aaaactcatc gagcatcaag tgaaactgca atttattcat
     4201 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     4261 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     4321 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     4381 accatgagtg acgactgaat ccggtgagaa tggcaagagt ttatgcattt ctttccagac
     4441 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcaccaa ccaaaccgtt
     4501 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgccgttaa aaggacaatt
     4561 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4621 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga tcgcagtggt
     4681 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     4741 ttccgtcagc cagtttagcc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     4801 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     4861 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     4921 ggaatttaat cgcggcttca agcaagacgt ttcccgttga atatggctca ttttagcttc
     4981 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     5041 atggtgaaag ttggaacctc ttacgtgccg atcacacatt tccccgaaaa gtgccacctg
     5101 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5161 cctttcgtc
//
