LOCUS       sequence-410557-        5969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 in astrocytes AAV Virus. Alias: AiP1349 - 
            pAAV-AiE2120m-minBG-SYFP2-WPRE-BGHpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5969)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, 
            Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith
            KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, 
            Levi BP, Ngai J, Ting JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 5969)
  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: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP1349 -
            pAAV-AiE2E120m-minBGpromoter-SYFP2-W...
FEATURES             Location/Qualifiers
     source          1..5969
                     /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      1981..1990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1987..1989,1990..1992,1993..2703)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2032..2053)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2293..2312)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2640..2661
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2796..2816)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3003..3114
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3232..3372
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3232..3361
                     /label=AAV2 ITR
     rep_origin      3447..3902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3534..3553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3744..3765
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3919..3938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4038..4060
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4098..4116)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4184..4288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4289..4357,4358..5149)
                     /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(4507..4526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5320..5908
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5809..5828
                     /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 gcgtcttgtt cttgttttga ggcagggtat tttataggca
      181 tagactttgc catgtaagcc aagctaagta tccctggacc ttcaagtgat ctgcctgcct
      241 ctgcctcctc caccatcatg aagctcacca ttgtcacagc agttttttac ctggatgctg
      301 gggaatctga actcaggttg tcacatttgc aaggctggta ctttaccaac tgaactacct
      361 ctttggccct actttttttg ttttcttgag aatgttctag aaagctctct gttaggtaat
      421 attttttttc atttgaagtt caatcttccc tttctgtttc taagactctc agagaaaatt
      481 aaacatttat tcttctatta agcttggaca agtactaaga ttaaaaaaaa aaaaagcaaa
      541 attagcattg ctttttcaag gcaaaagcaa agacagttcc tttatttaat aaccttcact
      601 catacaagaa ccaagctcat tagtcaagta ttggccacag aaaatgaaac tcttaagtct
      661 ccatgaaata atgtctttgg tcctatttag ccgagtgaag gatttagaca caaagcaaac
      721 agaaacttga aatctggtct tctcttctct ccagaatatt tggtctaaag taggtcagac
      781 cctgctgcaa ctaatcttaa cacttctttt gattctttca ctggctctac ttagaggagc
      841 ttagagaggg gaaaattcat ccaattagtt aaagaaaaac tgcaaatgag agaaaaggcc
      901 tcctgaattg tggtgcgggg tctctgtctc cagcgccaat gagttcatca acttgaaaat
      961 gattttcaaa agaatagctg tctatgaggt aatcattaat cccccggatg ctttagtggc
     1021 tgccctggct gccctcaacc atccattctc tctcctggag tctaagaagg atgctgacac
     1081 agctctggat gaagccaaga ccatgcctca agagaagaag agaaacctag aatgaccctc
     1141 agtccttcaa gacagccatc gcttcccaaa aaaaggccca gcaaggggtt tcaaaactct
     1201 agagttcaga agaaagttgc caggaggggc ctggattttg aaggtgtgta gaggaaggga
     1261 gtccactgaa ggtgttcata gtctacattg gtgtgtgcac atagcgtcta aatccaagcc
     1321 gttgcttttc cttggcctct cccatggatt cccacccgaa ccacagagtg ggacccttgt
     1381 ttccaagagt actttcgcca aaaaagtgca cagcttttca ccacttacgt gaaatgctgc
     1441 cagctaaata tccttctgaa tgatgtcttt gagacttccc aggcacttaa gctcctttct
     1501 aaaccatcac aggccaacaa agtctgtgag gttttaaggg atttttgctg gtcccgtggc
     1561 ttattctttg accacatcaa aatccaaggg attgctccag aaactgtgga tacagtttct
     1621 tttcttgttg ctatgaccaa agcaactaaa gaaggatttg tgtagctcac tgtttgaggg
     1681 tgtgatccat cgcaaagagg gaaggcgtgg ggacaggttc atgagacagc tattcacact
     1741 gtatgagagg ccctgaagca ggggagaaat gctggtatgc tggtgctcac ttttctttct
     1801 ttctttcttt ctttctttct tttttttttt tttttttttt ttttgacacg gtcttactat
     1861 gaagctctgc ttgtcttggt tctcccagag ctcgggctgg gcataaaagt cagggcagag
     1921 ccatctattg cttacatttg cttctgggat ccagatcttt cgaagctagc gctaccggtc
     1981 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2041 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2101 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     2161 cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac
     2221 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2281 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2341 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2401 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     2461 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     2521 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2581 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     2641 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2701 aagtaagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac
     2761 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2821 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2881 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     2941 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc
     3001 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3061 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3121 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3181 ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc
     3241 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3301 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3361 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3421 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3481 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     3541 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3601 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3661 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3721 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3781 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3841 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3901 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3961 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4021 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4081 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4141 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4201 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4261 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4321 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4381 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4441 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4501 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4561 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4621 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4681 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4741 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4801 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4861 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4921 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4981 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5041 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5101 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5161 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5221 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5341 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5401 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5461 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5521 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5581 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5641 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5701 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5761 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5821 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5881 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5941 ggccttttgc tggccttttg ctcacatgt
//
