LOCUS       sequence-233708-        6668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsRNA expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6668)
  TITLE     Petr Svoboda lab unpublished
REFERENCE   2  (bases 1 to 6668)
  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: CAG-EGFP-MosIR
FEATURES             Location/Qualifiers
     source          1..6668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1733..1742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1739..1741,1742..1744,1745..2458)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1739..1741,1742..1744,1745..2455)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1784..1805)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2045..2064)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2392..2413
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3690..3709)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3755..3810
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3809..3828)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4171..4187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4171..4187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4184..4206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4195..4211
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4219..4225,4226..4243,4244..4249))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4264..4285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4344..4539
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4390..4525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4545..4679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4582..4601)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4636..4655
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4747..4764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4918..5506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4998..5017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5677..6468,6469..6537))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6300..6319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6538..6642)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1801 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1861 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1921 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1981 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2041 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2101 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2161 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2221 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2281 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2341 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2401 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaga
     2461 attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg aattcggctt
     2521 agggtgattc caaaagagta gatgtcagct ttgggcgtgg caatctctcc tttcaggatc
     2581 tccggagctt ggtgcgtgta cgtgccccct atgtggtgag gggacgcctg ccggcaccgc
     2641 agatcctgca gcttctggga gcagccgaag tcactgatct tacaaacgtc ttgctcactg
     2701 atcaaaatgt tcgctggctt caggtccaag tgcaaaatgc tttgtgagtg gagaaaaagc
     2761 aggccgttaa caacatctag ggaatacttg aggcacttcc ccaaactcag ttgttctctg
     2821 cagctgagag gctccggtga gcgggtggca ccgtagatga cttggtgtag agtcacgttg
     2881 cccccaaact ccatgattat ggtacctagg ctgttggagt cttcgggcgt gcgcgtgctg
     2941 gcagccacaa cccgaactat gttgtcgtgg cgtagtcttg caatgttcag ttcagcccag
     3001 aaactccgct gggatgcacg taggtccttg gtgcaagggt gattccaaaa gagtaaaact
     3061 ttggtaaacc agattattta tccatctacc ccagtctgtg tttacccccc tttcttgttt
     3121 acttgcttga tggtgcacca aggacctacg tgcatcccag cggagtttct gggctgaact
     3181 gaacattgca agactacgcc acgacaacat agttcgggtt gtggctgcca gcacgcgcac
     3241 gcccgaagac tccaacagcc taggtaccat aatcatggag tttgggggca acgtgactct
     3301 acaccaagtc atctacggtg ccacccgctc accggagcct ctcagctgca gagaacaacc
     3361 gagtttgggg aagtgcctca agtattccct agatgttgtt aacggcctgc tttttctcca
     3421 ctcacaaagc attttgcact tggacctgaa gccagcgaac attttgatca gtgagcaaga
     3481 cgtttgtaag atcagtgact tcggctgctc ccagaagctg caggatctgc ggtgccggca
     3541 ggcgtcccct caccacatag ggggcacgta cacgcaccaa gctccggaga tcctgaaagg
     3601 agagattgcc acgcccaaag ctgacatcta ctcttttgga atcaccctaa gccgaattcc
     3661 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     3721 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     3781 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     3841 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     3901 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     3961 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     4021 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     4081 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
     4141 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4201 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4261 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4321 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     4381 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4441 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4501 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     4561 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4621 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     4681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6661 gccacctg
//
