LOCUS       sequence-390940-        5103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes dual cistronic cytosolic 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 5103)
  TITLE     plasmids optimized for C. elegans expression
REFERENCE   2  (bases 1 to 5103)
  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: pHJL2
FEATURES             Location/Qualifiers
     source          1..5103
                     /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      2850..3173
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(3224..3240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3224..3240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(3331..3919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3411..3430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4090..4905)
                     /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     4813..4832
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4906..5008)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     5076..5094
                     /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 caagtaagct gtctcatcct actttcacct agttaactgc ttgtcttaaa atctatgctt
     1741 ctctttagta tctaaaattt tcctagaagc ttacaagtat ataaatggtc tcttctcaat
     1801 aaaggttgta tatttattca tcttattgaa tctgccattt cctcgttttt gcgagtttat
     1861 ataccttcca attttctttc tattgtattt tcaacttcta attttaattc agggaaactg
     1921 cttcaacgca tcatggtttc caagggagag gctgttatca aggaattcat gacttacgaa
     1981 cttatcaagg agtttatgcg ctttaaggtt cacatggaag gatctatgaa cggacacgaa
     2041 ttcgaaatcg aaggagaagg agaaggacgt ccatacgagg gaactcaaac tgctaagctt
     2101 aaggtaagtt tatctaaaaa gtttttcatt caaaatgtgt aaaaattcat ttaaaataac
     2161 ccaaaaatca ttaatcctcg atattttcag gttactaaag gaggaccact tccattctct
     2221 tgggatatcc tttctccaca attcatgtac ggatctcgcg ctttcatcaa gcacccagct
     2281 gatatcccag attactacaa gcaatctttc ccagaaggat tcaaatggga gcgtgttatg
     2341 aacttcgaag atggaggagc tgttaccgtt acccaagata cctcccttga ggatggaacc
     2401 cttatctaca aggtattttc ctgcattttt caactgggaa aatgaaagaa aatcgataat
     2461 ttcaggttaa gcttcgtgga actaatttcc caccagatgg accagttatg caaaagaaga
     2521 ctatgggatg ggaggcttct accgagcgcc tttacccaga ggatggagtc cttaagggag
     2581 atatcaagat ggctcttcgt cttaaggatg gaggacgtta ccttgctgat ttcaaggtaa
     2641 gttttctaga gacctttact aactaactag gtctcataga ttttcagact acttacaagg
     2701 ctaagaagcc agttcaaatg ccgggagctt acaacgttga tcgtaagctt gatatcacct
     2761 ctcacaacga ggattacacc gttgttgagc aatacgagcg ttctgaggga cgtcactcta
     2821 ccggaggaat ggatgagctt tacaagtaaa tgcaaaatcc tttcaagcat tcccttcttc
     2881 tctatcactc ttctttcttt ttgtcaaaaa attctctcgc taatttattt gcttttttaa
     2941 tgttattatt ttatgacttt ttatagtcac tgaaaagttt gcatctgagt gaagtgaatg
     3001 ctatcaaaat gtgattctgt ctgatgtact ttcacaatct ctcttcaatt ccattttgaa
     3061 gtgctttaaa cccgaaaggt tgagaaaaat gcgagcgctc aaatatttgt attgtgttcg
     3121 ttgagtgacc caacaaaaag aggaaacttt attgtgccgc caagaaaaaa gtctcatacg
     3181 tatgatcggt ccacgatcag ctagattatc tagtcagctt gatgtcatag ctgtttcctg
     3241 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccagc aaaaggccag
     3301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgt cgcttaccgg
     3481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4081 gtctgacagt tagaaaaact catcgagcat caagtgaaac tgcaatttat tcatatcagg
     4141 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     4201 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     4261 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     4321 agtgacgact gaatccggtg agaatggcaa gagtttatgc atttctttcc agacttgttc
     4381 aacaggccag ccattacgct cgtcatcaaa atcactcgca ccaaccaaac cgttattcat
     4441 tcgtgattgc gcctgagcga gacgaaatac gcgatcgccg ttaaaaggac aattacaaac
     4501 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     4561 atcaggatat tcttctaata cctggaatgc tgttttccct gggatcgcag tggtgagtaa
     4621 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     4681 cagccagttt agcctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     4741 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     4801 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     4861 taatcgcggc ttcaagcaag acgtttcccg ttgaatatgg ctcattttag cttccttagc
     4921 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     4981 aaagttggaa cctcttacgt gccgatcaca catttccccg aaaagtgcca cctgacgtct
     5041 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5101 gtc
//
