LOCUS       sequence-292482-        6572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GoF mutant murine PCSK9 to be used for rAAV8-mediated gene
            transfer, hypercholesterolemia and atherosclerosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  AUTHORS   Bjorklund MM, Hollensen AK, Hagensen MK, Dagnaes-Hansen F, 
            Christoffersen C, Mikkelsen JG, Bentzon JF
  TITLE     Induction of atherosclerosis in mice and hamsters without germline 
            genetic engineering.
  JOURNAL   Circ Res. 2014 May 23;114(11):1684-9. doi: 
            10.1161/CIRCRESAHA.114.302937. Epub 2014 Mar 27.
  PUBMED    24677271
REFERENCE   2  (bases 1 to 6572)
  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"; doi: 
            "10.1161/CIRCRESAHA.114.302937"; journalName: "Circ Res"; date: 
            "2014-05-23- 23"; volume: "114"; issue: "11"; pages: "1684-9"
COMMENT     Sequence Label: pAAV/D377Y-mPCSK9
FEATURES             Location/Qualifiers
     source          1..6572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             2011..2022
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3509..3733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3835..3975
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3835..3964
                     /label=AAV2 ITR
     rep_origin      4050..4505
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4137..4156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4347..4368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4522..4541)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4641..4663
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4701..4719)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4787..4891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4892..4960,4961..5752)
                     /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(5110..5129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5923..6511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6412..6431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctgcag gctcagaggc acacaggagt ttctgggctc
      181 accctgcccc cttccaaccc ctcagttccc atcctccagc agctgtttgt gtgctgcctc
      241 tgaagtccac actgaacaaa cttcagccta ctcatgtccc taaaatgggc aaacattgca
      301 agcagcaaac agcaaacaca cagccctccc tgcctgctga ccttggagct ggggcagagg
      361 tcagagacct ctctgggccc atgccacctc caacatccac tcgacccctt ggaatttcgg
      421 tggagaggag cagaggttgt cctggcgtgg tttaggtagt gtgagagggt ccgggttcaa
      481 aaccacttgc tgggtgggga gtcgtcagta agtggctatg ccccgacccc gaagcctgtt
      541 tccccatctg tacaatggaa atgataaaga cgcccatctg atagggtttt tgtggcaaat
      601 aaacatttgg tttttttgtt ttgttttgtt ttgttttttg agatggaggt ttgctctgtc
      661 gcccaggctg gagtgcagtg acacaatctc atctcaccac aaccttcccc tgcctcagcc
      721 tcccaagtag ctgggattac aagcatgtgc caccacacct ggctaatttt ctatttttag
      781 tagagacggg tttctccatg ttggtcagcc tcagcctccc aagtaactgg gattacaggc
      841 ctgtgccacc acacccggct aattttttct atttttgaca gggacggggt ttcaccatgt
      901 tggtcaggct cctctagagg tacccgggga tcttgctacc agtggaacag ccactaagga
      961 ttctgcagtg agagcagagg gccagctaag tggtactctc ccagagactg tctgactcac
     1021 gccaccccct ccaccttgga cacaggacgc tgtggtttct gagccaggta caatgactcc
     1081 tttcggtaag tgcagtggaa gctgtacact gcccaggcaa agcgtccggg cagcgtaggc
     1141 gggcgactca gatcccagcc agtggactta gcccctgttt gctcctccga taactggggt
     1201 gaccttggtt aatattcacc agcagcctcc cccgttgccc ctctggatcc actgcttaaa
     1261 tacggacgag gacagggccc tgtctcctca gcttcaggca ccaccactga cctgggacag
     1321 tgaatgatcc ccctgatctg cggcctcgac ggtatcgata agcttgatat cgaattctag
     1381 tcgtcgacca ctttcacaat ctgcggcacc ggtaccatgg gcacccactg ctctgcgtgg
     1441 ctgcggtggc cgctgttgcc gctgttgccg ccgctgctgc tgctgttgct gctactgtgc
     1501 cccaccggcg ctggtgccca ggacgaggat ggagattatg aagagctgat gctcgccctc
     1561 ccgtcccagg aggatggcct ggctgatgag gccgcacatg tggccaccgc caccttccgc
     1621 cgttgctcca aggaggcctg gaggctgcca ggaacctaca ttgtggtgct gatggaggag
     1681 acccagaggc tacagattga acaaactgcc caccgcctgc agacccgggc tgcccgccgg
     1741 ggctatgtca tcaaggttct acatatcttt tatgacctct tccctggctt cttggtgaag
     1801 atgagcagtg acctgttggg cctggccctg aagttgcccc atgtggagta cattgaggaa
     1861 gactcctttg tcttcgccca gagcatccca tggaacctgg agcgaattat cccagcatgg
     1921 caccagacag aggaagaccg ctcccctgat ggaagcagcc aggtggaggt gtatctctta
     1981 gataccagca tccagggtgc ccatcgggag attgagggca gggtcaccat caccgacttc
     2041 aacagcgtgc cggaggagga tgggacacgc ttccacagac aggcgagcaa gtgtgacagc
     2101 cacggcaccc acctggcagg tgtggtcagc ggccgggatg ctggtgtggc caagggcacc
     2161 agcctgcaca gcctgcgtgt gctcaactgt caagggaagg gcacagtcag cggcaccctc
     2221 ataggcctgg agtttattcg gaagagtcag ctaatccagc cctcggggcc actcgtggtt
     2281 ctgctgcccc tggccggtgg gtatagccgc atcctcaacg ctgcctgccg gcacctggcg
     2341 aggactgggg tggtgctggt tgcagcagct gggaacttcc gggacgacgc ctgcctctac
     2401 tccccagctt ctgctccaga ggtcatcaca gtcggggcca cgaatgccca ggaccagcca
     2461 gttaccttgg ggactttggg gactaatttt ggacgctgtg tggatctctt tgcccccggg
     2521 aaggacatca tcggagcgtc cagttactgc agcacatgct tcatgtcaca gagtgggacc
     2581 tcacaggctg ctgcccacgt ggccggcatt gtggctcgga tgctgagccg ggagcccaca
     2641 cttaccctgg ccgagctgcg gcagaggctg atccacttct ctaccaaaga cgtcatcaac
     2701 atggcctggt tccctgagga ccagcaggtg ctgaccccca acctggtggc cacactgccc
     2761 cccagcaccc atgagacagg cgggcagctg ctctgtagga cggtgtggtc ggcacactcg
     2821 gggcccactc gaacagctac agctacagcc cgctgtgccc cagaagagga gctgctgagc
     2881 tgctccagct tctccaggag cgggaggcgt cgtggtgatt ggattgaggc cataggaggc
     2941 cagcaggtct gcaaggccct caatgcattt gggggtgagg gtgtctatgc cgtcgcgaga
     3001 tgctgcctgg ttccccgtgc caactgcagc atccacaaca cccctgcagc cagagctggc
     3061 ctggagaccc atgtccactg ccaccagaag gaccatgttc tcacaggctg cagcttccat
     3121 tgggaagtgg aagaccttag tgtccggagg cagcctgcgc tgaggtccag acgtcagcct
     3181 ggccagtgcg ttggccacca ggcggccagt gtctatgctt cctgctgcca tgccccaggg
     3241 ctggaatgca aaatcaagga gcatgggatc tcaggtcctt cagagcaggt cactgtggcc
     3301 tgcgaagcag gatggaccct gactggatgc aatgtgctcc ctggggcatc cctcactctg
     3361 ggagcctaca gcgtggacaa cctgtgtgtg gcaagagtcc atgacactgc cagagcagac
     3421 aggaccagtg gagaagccac agtagctgct gccatctgct gccggagccg gccttcagca
     3481 aaggcctcct gggttcagtg aggccggcct gtgccttcta gttgccagcc atctgttgtt
     3541 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3601 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3661 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3721 gtgggctcta tggcgcgccg cgatcgccca atacgcaaac cgcctctccc cgcgcggcgg
     3781 gtgtggtggt tacgcgcgct agccgaggta accacgtgcg gaccgagcgg ccgcaggaac
     3841 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3901 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3961 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4021 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4081 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     4141 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4201 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4261 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4321 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4381 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     4441 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4501 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4561 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4621 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4681 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4741 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4861 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4921 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4981 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5041 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5101 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5161 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5221 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5281 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5341 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5401 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5461 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5521 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5581 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5641 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5701 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5761 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5821 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6061 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6541 cctggccttt tgctggcctt ttgctcacat gt
//
