LOCUS       sequence-325811-        5391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing EGFP AAV Virus. Alias: AiP981 - 
            pAAV-AiE2004m-minBG-EGFP-WPRE-HGHpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5391)
  AUTHORS   Graybuck LT, Daigle TL, Sedeno-Cortes AE, Walker M, Kalmbach B, Lenz
            GH, Morin E, Nguyen TN, Garren E, Bendrick JL, Kim TK, Zhou T, 
            Mortrud M, Yao S, Siverts LA, Larsen R, Gore BB, Szelenyi ER, Trader
            C, Balaram P, van Velthoven CTJ, Chiang M, Mich JK, Dee N, Goldy J, 
            Cetin AH, Smith K, Way SW, Esposito L, Yao Z, Gradinaru V, Sunkin 
            SM, Lein E, Levi BP, Ting JT, Zeng H, Tasic B
  TITLE     Enhancer viruses for combinatorial cell-subclass-specific labeling.
  JOURNAL   Neuron. 2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011.
  PUBMED    33789083
REFERENCE   2  (bases 1 to 5391)
  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: "Neuron. 
            2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011."
COMMENT     Sequence Label: AiP981-pAAV-mscRE4-minBGpromoter-EGFP-WPRE-hGHpA
FEATURES             Location/Qualifiers
     source          1..5391
                     /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      768..777
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(774..776,777..779,780..1493)
                     /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(774..776,777..779,780..1490)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(819..840)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1080..1099)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1427..1448
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1518..2106
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1571..1591)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1989..2000)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2138..2614
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2337..2356)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2654..2794
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2654..2783
                     /label=AAV2 ITR
     rep_origin      2869..3324
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2956..2975)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3166..3187
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3341..3360)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3460..3482
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3520..3538)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3606..3710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3711..3779,3780..4571)
                     /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(3929..3948)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4742..5330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5231..5250
                     /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 gcgtataagc cttgggggca atcaaactat tacattgagt
      181 ccttggattt gctacaaatt acattttaaa tgcaatcatt ttataaaagc ttcaacactc
      241 acacttggaa gcgttaccct gttgaatatc actgactcac taacttgcat tgccatgcta
      301 acttgctttc agagagatct cagaacacat catcttctgc tatttcaata catgcacatt
      361 aatttcctat caacgtgtgc tgatcaggaa ctctgtaatc tggcaccggt gtttattttt
      421 attcctgtct attcctgttg gctcacgaaa agattgtttg agcaagtgtt ttatggtgag
      481 ttgtatcata tgtacattga tttaatctgc ccacattcag ttctacaagc ggagccaaaa
      541 aaatagagac aagcataatt ttcattcaac atgagcccct caatgcaagc caagtacctc
      601 atctggtgct cagctaaagc aacagcaatc tgttccaccc tggagacaca actggccaca
      661 gaaaacttag tgaaaagagg caatgctatg cacaggacaa atgagctcgg gctgggcata
      721 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccgcc accatggtga
      781 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      841 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      901 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      961 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     1021 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1081 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1141 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1201 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1261 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1321 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1381 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1441 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagaattcg
     1501 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1561 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1621 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     1681 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     1741 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     1801 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1861 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     1921 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     1981 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2041 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2101 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     2161 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     2221 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     2281 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     2341 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     2401 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     2461 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     2521 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     2581 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     2641 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2701 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2761 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2821 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2881 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2941 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3001 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3061 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3121 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3181 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     3241 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3301 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3361 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3421 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3481 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3541 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3601 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3661 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3721 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3781 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3841 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3901 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3961 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4021 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4081 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4141 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4201 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4261 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4321 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4381 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4441 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4501 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4561 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4621 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4681 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4741 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4801 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4861 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4921 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4981 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5041 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5101 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5161 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5221 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5281 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5341 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
