LOCUS       sequence-360689-        6312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV genome with a CAG driven Cre recombinase with mR122 target 
            sequence repeats to reduce transgene expression in hepatocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6312)
  AUTHORS   Krolak T, Chan KY, Kaplan L, Huang Q, Wu J, Zheng Q, Kozareva V, 
            Beddow T, Tobey IG, Pacouret S, Chen AT, Chan YA, Ryvkin D, Gu C, 
            Deverman BE
  TITLE     A High-Efficiency AAV for Endothelial Cell Transduction Throughout 
            the Central Nervous System.
  JOURNAL   Nat Cardiovasc Res. 2022 Apr;1(4):389-400. doi: 
            10.1038/s44161-022-00046-4. Epub 2022 Apr 13.
  PUBMED    35571675
REFERENCE   2  (bases 1 to 6312)
  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: 
            "10.1038/s44161-022-00046-4"; journalName: "Nat Cardiovasc Res"; 
            date: "2022-04"; volume: "1"; issue: "4"; pages: "389-400"
COMMENT     Sequence Label: pAAV-CAG-Cre-3xmiR122-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        210..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..793
                     /label=chicken beta-actin promoter
     primer_bind     1123..1145
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1207..1226
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1281..1301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2439..3027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2492..2512)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2910..2921)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3059..3535
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3258..3277)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3575..3715
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3575..3704
                     /label=AAV2 ITR
     rep_origin      3790..4245
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3877..3896)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4087..4108
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4262..4281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4381..4403
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4441..4459)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4527..4631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4632..4700,4701..5492)
                     /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(4850..4869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5663..6251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6152..6171
                     /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 gcgtaacact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      601 ttttgtgcag cgatgggggc gggggggggg ggggggsngc gcgccrggsg gggsggggsg
      661 gggsgrgggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      721 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      781 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      841 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      901 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
      961 gtggctgcgt gaaagccttg aggggctccg ggagcgccgg caggaaggaa atgggcgggg
     1021 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1081 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1141 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1201 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgcggccca
     1261 acggtaccgg atccaccatg cccaagaaga agaggaaggt gtccaatctc ctgactgttc
     1321 accagaacct ccctgcgctg ccagtagatg ccactagcga tgaggtcagg aaaaatctca
     1381 tggatatgtt tagggataga caggcgtttt ctgaacacac ctggaaaatg ctgcttagcg
     1441 tgtgccgatc ctgggcagcc tggtgtaagc tgaacaatcg caaatggttc cccgccgagc
     1501 cggaggacgt gcgcgattac ctgctgtatc tccaggcaag agggctggct gtcaagacta
     1561 tccagcagca cttgggccaa ctgaatatgc tgcatcgacg cagcgggctc ccccggccta
     1621 gcgattcaaa cgcagtctcc cttgttatga ggagaattag aaaggaaaac gtagatgcgg
     1681 gtgagagggc taagcaggct ctcgcttttg agcggactga tttcgaccag gtcagatccc
     1741 tgatggagaa cagcgatcgg tgccaggaca tcaggaacct cgcatttctg ggaattgcat
     1801 ataacacact tctgcgcata gctgagatcg cccggatcag agtgaaagac atcagtcgaa
     1861 cggacggcgg ccggatgctt attcatattg gacgcacaaa gacattggtc agcaccgctg
     1921 gcgttgaaaa ggccttgtcc ctgggcgtaa cgaagctggt ggaaagatgg atctcagtgt
     1981 ccggcgtggc tgacgaccct aataattact tgttctgtcg agtgagaaaa aacggagtcg
     2041 ccgcgccctc tgccaccagc caattgagta cacgggccct tgaagggatc tttgaggcaa
     2101 cccaccgact catatacgga gccaaggatg acagtggcca gaggtatctc gcctggtcag
     2161 gtcattctgc tagggtgggg gccgcacgag acatggcgcg ggcaggagtc tccataccag
     2221 agattatgca agctggaggt tggacaaatg tgaacatcgt tatgaactat atccgcaatc
     2281 ttgactctga aaccggggcc atggtgagac tgctcgaaga tggtgactga attcaccggt
     2341 caaacaccat tgtcacactc catactatca aacaccattg tcacactcca gatcaacaaa
     2401 caccattgtc acactccacg ccggcgaagc ttatcgataa tcaacctctg gattacaaaa
     2461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2821 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     2881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3001 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg
     3061 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     3121 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     3181 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     3241 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     3301 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     3361 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     3421 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     3481 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3541 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     3601 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3661 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3721 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3781 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3841 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     3901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4081 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     4141 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     4201 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4261 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4321 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4381 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4441 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4501 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4621 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4741 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4801 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4921 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5041 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5161 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5221 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5461 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5521 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6301 ttgctcacat gt
//
