LOCUS       sequence-243950-        6618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  siRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6618)
  TITLE     Oka Lab plasmids
REFERENCE   2  (bases 1 to 6618)
  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: AV5-siRNA-GFP
FEATURES             Location/Qualifiers
     source          1..6618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        109..421
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     340..360
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     437..453
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     438..454
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(496..529)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        645..948
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        949..1152
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1103..1123
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1191..1200
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1197..1199,1200..1202,1203..1916)
                     /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(1197..1199,1200..1202,1203..1913)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1242..1263)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1503..1522)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1850..1871
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1930..2518
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1983..2003)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2401..2412)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2526..3148
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2725..2744)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3245..3374
                     /label=AAV2 ITR
     promoter        complement(3445..3463)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3446..3463)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3481..3497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3481..3497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3494..3516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3505..3521
                     /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(3529..3535,3536..3553,3554..3559))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3574..3595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3883..4471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3963..3982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4642..5433,5434..5502))
                     /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     5265..5284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5503..5607)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5685..6140)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5822..5843)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6034..6053
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6266..6288
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6280..6297
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6281..6297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6304..6323
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6304..6322
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tttgcgacat tttgcgacac catgtggtca cgctgggtat ttaagcccga gtgagcacgc
       61 agggtctcca tacgcgccgt cgacgagctc agatctcccc gctctagaga tccgacgcgc
      121 catctctagg cccgcgccgg ccccctcgca cagacttgtg ggagaagctc ggctactccc
      181 ctgccccggt taatttgcat ataatatttc ctagtaacta tagaggctta atgtgcgata
      241 aaagacagat aatctgttct ttttaatact agctacattt tacatgatag gcttggattt
      301 ctataagaga tacaaatact aaattattat tttaaaaaac agcacaaaag gaaactcacc
      361 ctaactgtaa agtaattgtg tgttttgaga ctataaatat cccttggaga aaagccttgt
      421 taacgcgcgg tgaccctcga ggtcgacggt atcgataagc tcgcttcacg agattccagc
      481 aggtcgaggg acctaataac ttcgtatagc atacattata cgaagttata ttaagggttc
      541 caagcttaag cggccgcgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
      601 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      661 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      721 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      781 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      841 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      901 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      961 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1021 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1081 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1141 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg
     1201 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1261 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1321 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1381 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1441 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1501 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1561 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1621 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1681 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1741 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1801 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1861 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaggaat
     1921 tctatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1981 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2041 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2101 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2161 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2221 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2281 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2341 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2401 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2461 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     2521 cgatagggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     2581 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     2641 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     2701 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     2761 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     2821 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     2881 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     2941 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     3001 ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat aggctacctg
     3061 gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc tcatgcgttg
     3121 ggtccactca gtagatgcct gttgaattgg gctgcagacg cgtgtctaga ggagcatgcg
     3181 cgtcggcgcg ttctagagca tggctacgta gataagtagc atggcgggtt aatcattaac
     3241 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3301 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3361 gagcgagcgc gcagggttcg aaatcgataa gcttggatcg atccggagag ctcgtcgacg
     3421 gcgcgccatg catagcttga gtattctata gtgtcaccta aatagcttgg cgtaatcatg
     3481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt
     5641 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     5701 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5761 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5821 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5881 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5941 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6001 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6061 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     6121 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6181 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6241 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     6301 ttgtaatacg actcactata gggcgaattg ggcccggcgc gccgacgcgc atgctcctct
     6361 agctagactc gatcgaggaa ttcggtaccc cctgcgcgct cgctcgctca ctgaggccgc
     6421 ccgggcaaag cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     6481 tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag
     6541 ccatgctcta gagtggagtc gtgacgtgaa ttacgtcata gggttaggga ggtcctgtat
     6601 tagaggtcac gtgagtgt
//
