LOCUS       sequence-470447-        4983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0480m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4983)
  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 4983)
  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: AiP12259 - pAAV-AiE0480m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   354..483
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(545..1133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(625..644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1304..2095,2096..2164))
                     /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     1927..1946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2165..2269)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2337..2355
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2393..2415)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2515..2534
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2551..3006)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2688..2709)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2900..2919
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3081..3221
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3092..3221
                     /label=AAV2 ITR
     polyA_signal    3339..3450
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3637..3657
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3750..4460,4461..4463,4464..4466))
                     /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(3792..3813)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4141..4160
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4400..4421
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4463..4472
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 acattttttc agagtcataa ggcagtaatc acatttgtag aaaatgtgca gtaattatta
       61 atagctgttt ccaaattgaa ctaccaatta gaaacttcct agagaagccc aggatggttg
      121 ctatcttctc ctcagactga gaacaagcaa aatgtttcca gagaaaagcc aaggctaccc
      181 aggagtggga taataagcag gttaagaaaa tggtaataat agttattcat tatacccagg
      241 ttcaaatttg gcctctgggt caattaattt tcctttctgt gacaaaaata tttgagaaaa
      301 ccaacttaag gaagataaag tttctctgca gctcctggta cgcgtgcggc cgcaggaacc
      361 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
      421 accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
      481 aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1141 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1201 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1261 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1321 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     1381 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     1441 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1501 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1561 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1621 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     1681 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1741 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1801 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1861 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1921 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1981 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     2041 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     2101 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     2161 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2221 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     2281 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     2341 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2401 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2461 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     2521 cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat
     2581 tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa
     2641 tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca
     2701 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2761 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta
     2821 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2881 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa
     2941 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3001 gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag
     3061 aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     3121 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     3181 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac
     3241 gtgagatctc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc
     3301 caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt
     3361 aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa
     3421 caacagatgg ctggcaacta gaaggcacag tcgaagatct ctcgagccac ggaattgtca
     3481 gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc
     3541 gtggcaagaa ctaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca
     3601 atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag
     3661 ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atgatatcga
     3721 attcgcggcc gcggcgcgcc gtcgacttac ttgtacagct cgtccatgcc gagagtgatc
     3781 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     3841 ctcagcttgg actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     3901 atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg
     3961 cggatcttga agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg
     4021 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     4081 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     4141 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     4201 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     4261 acgccgtagc ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     4321 cagatcagct tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     4381 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     4441 aacagctcct cgcccttgct caccatggtg gcgaccggta gcgctagctt cgaaagatct
     4501 ggatcccaga agcaaatgta agcaatagat ggctctgccc tgacttttat gcccagcccg
     4561 agctcgctta cccttttcca ccactggatc tttcagtgaa acctgtcagg ctctgagatc
     4621 aatatttccg tttcttctcc tcagcctgtc tcctagatcc tcttatcagg attcttcaaa
     4681 cagggcctgt atctttctta gctcacacct gtcactgttt gcaatggagg taagagaaaa
     4741 ctgcaaaaat tgggcagaat attcagaggt acaattctca gctctacatg ttcacactgg
     4801 ggaatgtgtg tgtgtgtata catgtgtgca catgtgcata tgtgcataaa tgagaaacaa
     4861 ggaggaaggg tcctgacagg gaaccgtcgt tctgggtgac gtttagagca ggaagtccat
     4921 ttaagaatca tctgtttctt tcttgtctat ttcgggtaat gaaacagtga cccagagctg
     4981 tca
//
