LOCUS       sequence-333468-        6571 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses GoF mutant human 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 6571)
  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 6571)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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/D374Y-hPCSK9
FEATURES             Location/Qualifiers
     source          1..6571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1416..1425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2007..2018
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(3104..3115)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    3508..3732
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3834..3974
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3834..3963
                     /label=AAV2 ITR
     rep_origin      4049..4504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4136..4155)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4346..4367
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4521..4540)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4640..4662
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4700..4718)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4786..4890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4891..4959,4960..5751)
                     /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(5109..5128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5922..6510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6411..6430
                     /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 ggtgcgccac catgggcacc gtcagctcca
     1441 ggcggtcctg gtggccgctg ccactgctgc tgctgctgct gctgctcctg ggtcccgcgg
     1501 gcgcccgtgc gcaggaggac gaggacggcg actacgagga gctggtgcta gccttgcgtt
     1561 ccgaggagga cggcctggcc gaagcacccg agcacggaac cacagccacc ttccaccgct
     1621 gcgccaagga tccgtggagg ttgcctggca cctacgtggt ggtgctgaag gaggagaccc
     1681 acctctcgca gtcagagcgc actgcccgcc gcctgcaggc ccaggctgcc cgccggggat
     1741 acctcaccaa gatcctgcat gtcttccatg gccttcttcc tggcttcctg gtgaagatga
     1801 gtggcgacct gctggagctg gccttgaagt tgccccatgt cgactacatc gaggaggact
     1861 cctctgtctt tgcccagagc atcccgtgga acctggagcg gattacccct ccacggtacc
     1921 gggcggatga ataccagccc cccgacggag gcagcctggt ggaggtgtat ctcctagaca
     1981 ccagcataca gagtgaccac cgggaaatcg agggcagggt catggtcacc gacttcgaga
     2041 atgtgcccga ggaggacggg acccgcttcc acagacaggc cagcaagtgt gacagtcatg
     2101 gcacccacct ggcaggggtg gtcagcggcc gggatgccgg cgtggccaag ggtgccagca
     2161 tgcgcagcct gcgcgtgctc aactgccaag ggaagggcac ggttagcggc accctcatag
     2221 gcctggagtt tattcggaaa agccagctgg tccagcctgt ggggccactg gtggtgctgc
     2281 tgcccctggc gggtgggtac agccgcgtcc tcaacgccgc ctgccagcgc ctggcgaggg
     2341 ctggggtcgt gctggtcacc gctgccggca acttccggga cgatgcctgc ctctactccc
     2401 cagcctcagc tcccgaggtc atcacagttg gggccaccaa tgcccaggac cagccggtga
     2461 ccctggggac tttggggacc aactttggcc gctgtgtgga cctctttgcc ccaggggagg
     2521 acatcattgg tgcctccagc tactgcagca cctgctttgt gtcacagagt gggacatcac
     2581 aggctgctgc ccacgtggct ggcattgcag ccatgatgct gtctgccgag ccggagctca
     2641 ccctggccga gttgaggcag agactgatcc acttctctgc caaagatgtc atcaatgagg
     2701 cctggttccc tgaggaccag cgggtactga cccccaacct ggtggccgcc ctgcccccca
     2761 gcacccatgg ggcaggttgg cagctgtttt gcaggactgt gtggtcagca cactcggggc
     2821 ctacacggat ggccacagcc atcgcccgct gcgccccaga tgaggagctg ctgagctgct
     2881 ccagtttctc caggagtggg aagcggcggg gcgagcgcat ggaggcccaa gggggcaagc
     2941 tggtctgccg ggcccacaac gcttttgggg gtgagggtgt ctacgccatt gccaggtgct
     3001 gcctgctacc ccaggccaac tgcagcgtcc acacagctcc accagctgag gccagcatgg
     3061 ggacccgtgt ccactgccac caacagggcc acgtcctcac aggctgcagc tcccactggg
     3121 aggtggagga ccttggcacc cacaagccgc ctgtgctgag gccacgaggt cagcccaacc
     3181 agtgcgtggg ccacagggag gccagcatcc acgcttcctg ctgccatgcc ccaggtctgg
     3241 aatgcaaagt caaggagcat ggaatcccgg cccctcagga gcaggtgacc gtggcctgcg
     3301 aggagggctg gaccctgact ggctgcagtg ccctccctgg gacctcccac gtcctggggg
     3361 cctacgccgt agacaacacg tgtgtagtca ggagccggga cgtcagcact acaggcagca
     3421 ccagcgaaga ggccgtgaca gccgttgcca tctgctgccg gagccggcac ctggcgcagg
     3481 cctcccagga gctccagtga ggccggcctg tgccttctag ttgccagcca tctgttgttt
     3541 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3601 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3661 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3721 tgggctctat ggcgcgccgc gatcgcccaa tacgcaaacc gcctctcccc gcgcggcggg
     3781 tgtggtggtt acgcgcgcta gccgaggtaa ccacgtgcgg accgagcggc cgcaggaacc
     3841 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3901 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3961 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4021 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4081 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     4141 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4201 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4261 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4321 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4381 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4441 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4501 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4561 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4621 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4681 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4741 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4801 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4861 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4921 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4981 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5041 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5101 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5161 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5221 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5281 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5341 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5401 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5461 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5521 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5581 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5641 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5701 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5761 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5821 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5881 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5941 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6001 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6061 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6121 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6181 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6241 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6301 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6361 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6421 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6481 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6541 ctggcctttt gctggccttt tgctcacatg t
//
