LOCUS       sequence-239714-        6550 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 6550)
  TITLE     Petr Svoboda lab unpublished
REFERENCE   2  (bases 1 to 6550)
  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-Elavl2IR
FEATURES             Location/Qualifiers
     source          1..6550
                     /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(3572..3591)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3637..3692
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3691..3710)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4053..4069)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4053..4069)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4066..4088)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4077..4093
                     /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(4101..4107,4108..4125,4126..4131))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4146..4167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4226..4421
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4272..4407
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4427..4561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4464..4483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4518..4537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4629..4646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4800..5388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4880..4899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5559..6350,6351..6419))
                     /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     6182..6201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6420..6524)
                     /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 gggggggcgc gccrgggggg gggggggggg 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 ccaatgccct
     2521 ggctcgagat agctagcttc gcggccgcga ctctagaaaa ccctacaccc cttgatatgc
     2581 cagtgacctg gtcgacaaga atccgagaag taatgatgcg cccatactgt gaaaacagct
     2641 gttccagctc cttctgggtc atggtttttg gaagcccgct gacgtataag tttgcatctc
     2701 tgattgaggc tgagcttggg cgagcatagg aaacttttgt tgttttggtt tgaagtctca
     2761 atccattcag agtgttgata gctttctctg catccttggg gtcaatgtag ttcacaaatc
     2821 cgtaacccaa gctctgccct gttattttgt ctcttacaag cttacaggac tctatctcgc
     2881 caatgctccc aaaaagactc tttagttctt cctgtgtcat gttctgagga aggtagttga
     2941 ctattaggtt ggtcttgctg tcctctgtgt tcccggagtc aactggtgag gagcagttgt
     3001 tgtttacagt ggttggacca ttggctgtgt tattgcaagt tggcccatta gacagttgtg
     3061 tttccaggat ccggtccaac cactgtaaac aacaactgct cctcaccagt tgactccggg
     3121 aacacagagg acagcaagac caacctaata gtcaactacc ttcctcagaa catgacacag
     3181 gaagaactaa agagtctttt tgggagcatt ggcgagatag agtcctgtaa gcttgtaaga
     3241 gacaaaataa cagggcagag cttgggttac ggatttgtga actacattga ccccaaggat
     3301 gcagagaaag ctatcaacac tctgaatgga ttgagacttc aaaccaaaac aacaaaagtt
     3361 tcctatgctc gcccaagctc agcctcaatc agagatgcaa acttatacgt cagcgggctt
     3421 ccaaaaacca tgacccagaa ggagctggaa cagctgtttt cacagtatgg gcgcatcatt
     3481 acttctcgga ttcttgtcga ccaggtcact ggcatatcaa ggggtgtagg gttttctaga
     3541 gcggccgcgt tggtaccaac catggtgtag atctttttcc ctctgccaaa aattatgggg
     3601 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     3661 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     3721 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     3781 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     3841 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     3901 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     3961 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     4021 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4081 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4141 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4201 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     4261 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4321 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4381 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     4441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4561 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4681 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4741 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5221 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5461 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5521 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5581 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5641 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5701 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5761 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5821 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5881 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5941 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6001 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6061 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6121 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6181 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6241 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6301 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6361 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6421 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6481 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6541 gtgccacctg
//
