LOCUS       sequence-356206-        6311 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 6311)
  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 6311)
  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..6311
                     /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..792
                     /label=chicken beta-actin promoter
     primer_bind     1122..1144
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1206..1225
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1280..1300
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2438..3026
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2491..2511)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2909..2920)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3058..3534
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3257..3276)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3574..3714
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3574..3703
                     /label=AAV2 ITR
     rep_origin      3789..4244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3876..3895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4086..4107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4261..4280)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4380..4402
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4440..4458)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4526..4630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4631..4699,4700..5491)
                     /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(4849..4868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5662..6250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6151..6170
                     /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 ggggggcgcg cgccaggcgg ggcggggcgg
      661 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga
     1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa
     1261 cggtaccgga tccaccatgc ccaagaagaa gaggaaggtg tccaatctcc tgactgttca
     1321 ccagaacctc cctgcgctgc cagtagatgc cactagcgat gaggtcagga aaaatctcat
     1381 ggatatgttt agggatagac aggcgttttc tgaacacacc tggaaaatgc tgcttagcgt
     1441 gtgccgatcc tgggcagcct ggtgtaagct gaacaatcgc aaatggttcc ccgccgagcc
     1501 ggaggacgtg cgcgattacc tgctgtatct ccaggcaaga gggctggctg tcaagactat
     1561 ccagcagcac ttgggccaac tgaatatgct gcatcgacgc agcgggctcc cccggcctag
     1621 cgattcaaac gcagtctccc ttgttatgag gagaattaga aaggaaaacg tagatgcggg
     1681 tgagagggct aagcaggctc tcgcttttga gcggactgat ttcgaccagg tcagatccct
     1741 gatggagaac agcgatcggt gccaggacat caggaacctc gcatttctgg gaattgcata
     1801 taacacactt ctgcgcatag ctgagatcgc ccggatcaga gtgaaagaca tcagtcgaac
     1861 ggacggcggc cggatgctta ttcatattgg acgcacaaag acattggtca gcaccgctgg
     1921 cgttgaaaag gccttgtccc tgggcgtaac gaagctggtg gaaagatgga tctcagtgtc
     1981 cggcgtggct gacgacccta ataattactt gttctgtcga gtgagaaaaa acggagtcgc
     2041 cgcgccctct gccaccagcc aattgagtac acgggccctt gaagggatct ttgaggcaac
     2101 ccaccgactc atatacggag ccaaggatga cagtggccag aggtatctcg cctggtcagg
     2161 tcattctgct agggtggggg ccgcacgaga catggcgcgg gcaggagtct ccataccaga
     2221 gattatgcaa gctggaggtt ggacaaatgt gaacatcgtt atgaactata tccgcaatct
     2281 tgactctgaa accggggcca tggtgagact gctcgaagat ggtgactgaa ttcaccggtc
     2341 aaacaccatt gtcacactcc atactatcaa acaccattgt cacactccag atcaacaaac
     2401 accattgtca cactccacgc cggcgaagct tatcgataat caacctctgg attacaaaat
     2461 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2521 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2581 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2641 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2701 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2761 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2821 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     2881 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2941 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3001 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
     3061 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3121 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3181 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3241 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3301 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     3361 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3421 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3481 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     3541 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3601 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3661 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3721 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     3781 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3841 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     3901 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3961 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4021 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4081 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4141 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4201 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4261 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4321 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4381 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4441 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4501 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4561 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4621 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4681 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4741 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4801 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4861 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4921 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4981 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5041 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5101 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5161 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5221 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5281 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5341 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5401 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5461 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5521 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5581 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5641 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5701 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5761 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5821 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5881 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5941 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6001 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6061 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6301 tgctcacatg t
//
