LOCUS       sequence-389907-        6553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control vector expressing a U7-based antisense oligonucleotide 
            targeting CypA exon 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6553)
  AUTHORS   van der Wal E, Bergsma AJ, Pijnenburg JM, van der Ploeg AT, 
            Pijnappel WWMP
  TITLE     Antisense Oligonucleotides Promote Exon Inclusion and Correct the 
            Common c.-32-13T>G GAA Splicing Variant in Pompe Disease.
  JOURNAL   Mol Ther Nucleic Acids. 2017 Jun 16;7:90-100. doi: 
            10.1016/j.omtn.2017.03.001. Epub 2017 Mar 14.
  PUBMED    28624228
REFERENCE   2  (bases 1 to 6553)
  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"; journalName: "Mol Ther 
            Nucleic Acids."; date: "2017-06-16"; volume: "7:90-100. doi"; pages:
            " 10.1016/j.omtn.2017.03.001. Epub 2017 Mar 14"
COMMENT     Sequence Label: SF-U7snRNA-CypA-E4
FEATURES             Location/Qualifiers
     source          1..6553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1802..1919
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2397..2985
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2450..2470)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2868..2879)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3072..3305
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3377..3498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3414..3433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3468..3487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3538..3673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3693..3712)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3694..3712)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3722..3739)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3722..3738)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3731..3753)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3880..4335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3967..3986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4177..4198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4361..4465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4466..4534,4535..5326)
                     /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     complement(4684..4703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5497..6085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5986..6005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6239..6256
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6373..6394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6409..6414,6415..6432,6433..6439)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6447..6463
                     /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)."
     primer_bind     6452..6474
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6471..6487
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6471..6487
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6506..6526
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6508..6526
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta
     1921 tcgatcacga gactagcctc gagctagctg cagtaacaac ataggagctg tgattggctg
     1981 ttttcagcca atcagcactg actcatttgc atagccttta caagcggtca caaactcaag
     2041 aaacgagcgg ttttaatagt cttttagaat attgtttatc gaaccgaata aggaactgtg
     2101 ctttgtgatt cacatatcag tggaggggtg tggaaatggc accttgatct caccctcatc
     2161 gaaagtggag ttgatgtcct tccctggctc gctacagatg catgtaccct taccactcag
     2221 ttcaccaccc tgtcaacata gaatttttgg agcaggtttt ctgacttcgg tcggaaaacc
     2281 cctcccaatt tcactggtct acaatgaaag caaaacagtt ctcttccccg ctccccggtg
     2341 tgtgagaggg gctttgatcc ttctctggtt tcctaggaaa cgcgtatctg gtcgacaatc
     2401 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2461 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2521 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2581 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2641 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2701 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2761 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     2821 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2881 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2941 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc
     3001 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     3061 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     3121 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     3181 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     3241 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     3301 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     3361 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3421 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3481 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     3541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3601 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3661 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     3721 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     3781 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     3841 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     3901 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3961 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4021 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4081 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4141 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4201 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4261 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4321 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4381 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4441 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4501 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4561 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4621 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4681 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4741 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4801 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4861 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4921 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4981 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5041 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5101 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5161 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5221 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5281 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5341 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5401 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5461 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5521 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5581 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5641 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5701 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5761 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5821 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5881 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5941 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6001 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6061 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6121 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6181 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6241 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6301 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6361 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6421 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6481 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     6541 gctgcaagct taa
//
