LOCUS       sequence-421847-        5370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to restrict GFP expression in microglia..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5370)
  AUTHORS   Serrano C, Cananzi S, Shen T, Wang LL, Zhang CL
  TITLE     Simple and highly specific targeting of resident microglia with 
            adeno-associated virus.
  JOURNAL   iScience. 2024 Aug 12;27(9):110706. doi: 10.1016/j.isci.2024.110706.
            eCollection 2024 Sep 20.
  PUBMED    39297168
REFERENCE   2  (bases 1 to 5370)
  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: "iScience.";
            date: "2024-08-12"; volume: "27(9):110706. doi"; pages: " 
            10.1016/j.isci.2024.110706. eCollection 2024 Sep 20"
COMMENT     Sequence Label: pAAV-hIBA1a-GFP-miR124T
FEATURES             Location/Qualifiers
     source          1..5370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      163..618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(250..269)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     460..481
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(635..654)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     754..776
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(814..832)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        900..1004
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1005..1073,1074..1865)
                     /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(1223..1242)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2036..2624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2525..2544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2686..2815
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3302..3311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3308..3310,3311..3313,3314..4024)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3308..3310,3311..3313,3314..4024)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3353..3374)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3614..3633)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3961..3982
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4178..4766
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4231..4251)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4649..4660)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4802..5278
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5001..5020)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   join(5318..5370,1..88)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5318..5370,1..77)
                     /label=AAV2 ITR
ORIGIN
        1 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
       61 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
      121 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
      181 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
      241 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      301 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      361 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      421 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      481 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
      541 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
      601 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      661 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      721 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
      781 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
      841 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
      901 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      961 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1021 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1081 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1141 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1201 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1261 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1321 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1381 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1441 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     1501 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     1561 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     1621 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     1681 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     1741 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     1801 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     1861 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1921 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1981 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2041 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2101 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2161 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2221 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2281 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2341 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2401 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2461 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2521 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2581 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2641 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc
     2701 gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca
     2761 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
     2821 cgcacgcgaa ttcagccagc ccttccttgt cttcctatcc acagctgcct gcccagactc
     2881 atgaaacctg acaaaatgca aggtcttatc attacctgaa ccttggacct gttcaaaaat
     2941 actagttcct gagaataaat atccctggtg tcttcctgcc cttcctgcac acctccagtg
     3001 gcttatcaaa atatttgttt catgcgcaca ctgggctctc atttaagagg aatttgggag
     3061 aatgttattt tctaatctgc atttcacacc aggctccccc tccttcctgg ggtgctagtg
     3121 tcagcagaac ctgatgggga agtgaggtct gggaggcaga ggaggaagga atgaggggaa
     3181 aggggaagtt tgggaggaag gcttctgaga agactggtgg gagagaagga gagcctgcag
     3241 acagaggcct ccagcttggt ctgtctcccc acctctacca gcatctgctg agctaccggt
     3301 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     3361 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3421 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     3481 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3541 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3601 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3661 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3721 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3781 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3841 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3901 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3961 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     4021 caagtccgga ctcagatcct actagtagga tctcgaggga tcaagcttgt cgactggcat
     4081 tcaccgcgtg ccttacagtt ggcattcacc gcgtgcctta ctcgactggc attcaccgcg
     4141 tgccttacat tggcattcac cgcgtgcctt aatcgataat caacctctgg attacaaaat
     4201 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4261 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4321 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4381 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4441 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4501 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4561 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     4621 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4681 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4741 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgatc gagagatcta
     4801 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     4861 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     4921 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     4981 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     5041 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     5101 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     5161 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     5221 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     5281 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     5341 gccactccct ctctgcgcgc tcgctcgctc
//
