LOCUS       sequence-496013-        7366 bp    DNA     circular SYN 23-DEC-2025
DEFINITION  Microglia-targeted transgene expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7366)
  AUTHORS   Okada Y, Hosoi N, Matsuzaki Y, Fukai Y, Hiraga A, Nakai J, Nitta K, 
            Shinohara Y, Konno A, Hirai H
  TITLE     Development of microglia-targeting adeno-associated viral vectors as
            tools to study microglial behavior in vivo.
  JOURNAL   Commun Biol. 2022 Nov 11;5(1):1224. doi: 10.1038/s42003-022-04200-3.
  PUBMED    36369525
REFERENCE   2  (bases 1 to 7366)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2022-11-11"; volume: "5(1):1224. doi"; pages: " 
            10.1038/s42003-022-04200-3"
COMMENT     Sequence Label: pAAV/mIba1.GFP.WPRE.miR-9.T.miR-129-2-3p.T.SV40pA
FEATURES             Location/Qualifiers
     source          1..7366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      2678..2687
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2684..2686,2687..2689,2690..3403)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2684..2686,2687..2689,2690..3400)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2729..2750)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2990..3009)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3337..3358
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3419..4007
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3472..3492)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3890..3901)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4302..4321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4356..4375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4471..4600
                     /label=AAV2 ITR
     rep_origin      4971..5484
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5058..5077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5268..5289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5966..5985)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6085..6107
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6145..6163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6231..6335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6336..6404,6405..7196)
                     /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(6554..6573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg cgctcgctcg
      841 ctcactgagg ccgcccgggc aaagcccggg cggcctcagt gagcgagcga gcgcgcagag
      901 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
      961 acttatctac gtagccatgc tctagactcg agtactatag gatgcatcgt gaaaacctcg
     1021 tttccaccag gaactgaggt tgctgctgga ggaaatctcc catgaagagc tgttaactgg
     1081 ggaactggtt ggatccagct cctacaagct ctcctttaat ttccatacac atgccaagct
     1141 gtgaacacct gcacacacag tacaaataaa acagtagaaa tgaaatgaaa attaaaacaa
     1201 aaacagcata ctttctcctt tctacccaca tatctcctgc atccctgaga gcaatttctt
     1261 ggccatcttc cataatcatg ctattcaaac ctgccctcta agtacagagc tctgaatgga
     1321 gaccacggga gcagactttt catctgttgg ctccctgggc ctagcagtgt accctgcaaa
     1381 caggaaacct tcgctcatca tttatcacat gaatgtggct agagaactgt ccccaccaga
     1441 cactgagagc ctctcacagg gccagagggt ggctgcctca aaggcaggcg agaggctgca
     1501 gtcttctgca gagtggacag atactgcctg ccatacaagg ggacatagcc tggggcgggg
     1561 ggcacaaggg aaggggcctg agtgcattgg agtcacagcc ctccctcccc tggtgaccca
     1621 gtgactgagg ggaccaaggc tatccctggt atgaggggag ggcatttgca tgagttagac
     1681 ccctgacagc ccatggtggg gacaggaagt agctctgtct actgtctctt tcagtctcac
     1741 tttctgttcc tccaaagtgt ctcttcaaac tctcaaatga ctttgtagat tcagcctaag
     1801 atctcaaagc aggtgaggca ggaggctggc actcaagagc ctgacgaaca caaacccaag
     1861 tccttctggg acatgtctcc atgtgtcccc agtgctcctc tgtagcccct gagagaggtt
     1921 tctttccttt ccagggcgct aggctcagct caccccattc ctggagcagc ctgcagactt
     1981 catcctctct cttccatccc ggggaaagtc agccagtcct cctcagctgc ctgtcttaac
     2041 ctgcatcatg aagcctgagg agatttcaag taaaccctcc caagccccac ctcaggatct
     2101 ggggaaagcc actgtctacc gcatccttgg tttgagacag gttcttactg tgttggcctg
     2161 gctgggttcc aactcccagg atcctctagc ctcagcctct cttaggtgct agattgcagg
     2221 cctagggtat cacgctcggt ccctcagatc ctggtgcctt tccacacctc cgggagctga
     2281 tctaagtctt tctcccacgt acagtctgct tctttggagg agctgtgtgg agctaggtgt
     2341 gttacagggc tgtagctcca gccatctggg aggctgaggc aggatggcta cttcaattct
     2401 caagagcctg gataggagac cctgccccca ccccaccccc aaagtaattt ttcagcacat
     2461 tacttcttca tctcctctct caacccgttc tgccttctcc tggggtgctg gtgtcagcag
     2521 aagctgatgt ggaagtgatg cctgggagtt agcaagggaa tgagtggaaa ggggaagtgt
     2581 gagaacggtc ccagaagaga ctggggagct ggtggagaga ggacccagcg gacagactgc
     2641 cagcctaaga caaccagcgt ctgaggagcc accggtcgcc accatggtga gcaagggcga
     2701 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2761 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2821 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2881 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2941 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3001 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3061 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3121 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3181 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3241 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3301 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3361 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcatcgataa
     3421 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3481 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3541 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3601 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3661 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3721 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3781 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     3841 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3901 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3961 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gatgccgtat
     4021 acggtacctc atacagctag ataaccaaag cgattcatac agctagataa ccaaagcaat
     4081 tgtcatacag ctagataacc aaagtcactc atacagctag ataaccaaag gctagcatgc
     4141 tttttggggt aagggcttcg atatgctttt tggggtaagg gcttcttaag atgctttttg
     4201 gggtaagggc tttcacatgc tttttggggt aagggcttgg atcctctagt agagtcgagg
     4261 aattcacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4321 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4381 atcttatcat gtctgtcgac ccccaacgcg ttggagctct agagcatggc tacgtagata
     4441 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     4501 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4561 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg
     4621 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgattccgtt
     4681 gcaatggctg gcggtaatat tgttctggat attaccagca aggccgatag tttgagttct
     4741 tctactcagg caagtgatgt tattactaat caaagaagta ttgcgacaac ggttaatttg
     4801 cgtgatggac agactctttt actcggtggc ctcactgatt ataaaaacac ttctcaggat
     4861 tctggcgtac cgttcctgtc taaaatccct ttaatcggcc tcctgtttag ctcccgctct
     4921 gattctaacg aggaaagcac gttatacgtg ctcgtcaaag caaccatagt acgcgccctg
     4981 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5041 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5101 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5161 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5221 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5281 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5341 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5401 taacaaaata ttaacgctta caatttaaat atttgcttat acaatcttcc tgtttttggg
     5461 gcttttctga ttatcaaccg gggtacatat gattgacatg ctagttttac gattaccgtt
     5521 catcgattct cttgtttgct ccagactctc aggcaatgac ctgatagcct ttgtagagac
     5581 ctctcaaaaa tagctaccct ctccggcatg aatttatcag ctagaacggt tgaatatcat
     5641 attgatggtg atttgactgt ctccggcctt tctcacccgt ttgaatcttt acctacacat
     5701 tactcaggca ttgcatttaa aatatatgag ggttctaaaa atttttatcc ttgcgttgaa
     5761 ataaaggctt ctcccgcaaa agtattacag ggtcataatg tttttggtac aaccgattta
     5821 gctttatgct ctgaggcttt attgcttaat tttgctaatt ctttgccttg cctgtatgat
     5881 ttattggatg ttggaatcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5941 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6001 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6061 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6121 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     6181 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6541 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6721 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6841 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7081 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7201 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7261 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
