LOCUS       sequence-411044-        5161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for SYFP2 expression in brain astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5161)
  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 5161)
  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: AiP12214 - pAAV-ProB12-SYFP2-WPRE3-BGHpA (Alia...
FEATURES             Location/Qualifiers
     source          1..5161
                     /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      1173..1182
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1179..1181,1182..1184,1185..1895)
                     /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(1224..1245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1485..1504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1832..1853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1988..2008)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2195..2306
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2424..2564
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2424..2553
                     /label=AAV2 ITR
     rep_origin      2639..3094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2726..2745)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2936..2957
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3111..3130)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3230..3252
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3290..3308)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3376..3480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3481..3549,3550..4341)
                     /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(3699..3718)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4512..5100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5001..5020
                     /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 gcgtaccctc ctcaggggaa gaaacagtac attctctgaa
      181 gcctcctggt taaatggggg aagtatttgt attcctatca ccaagtagga gtccttcagt
      241 agatgaaagg ggtattttaa accgttgaag ctatcttggt ggcaatctag gatgttgaga
      301 cctcagagaa aatgtcgaaa ccttggaaaa tttattttga cgattagaaa ttgtttaata
      361 tctccttcca agattaatct atgattaaag ggcacttaag agacttttat aatttctaaa
      421 atattcaata gcatagatga tttggccatt acaaactgcc taagatttct cctcaggcag
      481 aagccaggcc acccacggag agacccagga actgggccaa gagcctaaac aaaagacttc
      541 ctttctccag ccccactttc cctgtttctt ttgtcatagg ttgttgtgaa ctaaaaaaat
      601 agctcagttt cacaaaagcg atctgggagg gaatcctgga ctcgcaaccc gactccgctc
      661 ggcgttgatt ggccccgcct cccagcagct caatagttac aggggagtgg caggtccctc
      721 ccggccacgt cttggacccg ccccccggcc gtgggaatcg ccgcctagca ggcggacaga
      781 ctggagaggt ggattaatga ccccagcaaa aagctggcat gtccaagaga tttaaaatat
      841 tttttttatg taacagcttt agccaaagct gttctagggt tcagaggaac tgccactcag
      901 tcggagtggc ggaggccgta gccccgcctc ctccccccta attgatagac tgacttcttg
      961 actaatggcc aatctctaag cagaggggag gaccttccta ttggccagag agtcagcttt
     1021 caaataaaag aggcggctct tccggcaaca aagaaaacaa ggcgagccta taaacaaagc
     1081 cacgtgggct tcaagtttcc acggacgcaa ggttgtgcaa cgcagtagtc acttattcgg
     1141 atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1201 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1261 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
     1321 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
     1381 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1441 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1501 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1561 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1621 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     1681 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1741 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     1801 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1861 ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc
     1921 gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1981 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2041 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg
     2101 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2161 gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct
     2221 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2281 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2341 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc
     2401 tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     2461 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     2521 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     2581 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     2641 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2701 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2761 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2821 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     2881 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2941 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     3001 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3061 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     3121 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3301 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3481 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3541 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3601 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3661 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3721 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3781 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3841 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3901 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3961 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4021 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4081 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4141 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4201 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4261 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4321 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4381 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4441 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4501 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4561 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4621 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4681 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4741 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4801 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4861 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4921 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4981 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5041 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5101 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5161 t
//
