LOCUS       sequence-466282-        4895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2335m 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 4895)
  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 4895)
  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: AiP1655 - pAAV-AiE2335m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..79
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(233..252)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     352..374
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(412..430)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        498..602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(603..671,672..1463)
                     /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(821..840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1634..2222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2123..2142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2284..2413
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3190..3199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3196..3198,3199..3201,3202..3912)
                     /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(3241..3262)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3502..3521)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3849..3870
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4005..4025)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4212..4323
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4441..4581
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4441..4570
                     /label=AAV2 ITR
     rep_origin      join(4656..4895,1..216)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4743..4762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
       61 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
      121 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
      181 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
      241 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
      301 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
      361 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
      421 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
      481 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      541 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      601 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      661 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      721 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      781 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      841 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      901 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      961 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1021 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1081 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1141 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1201 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1261 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1321 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1381 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1441 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1501 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1561 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1621 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1681 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1741 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1801 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1861 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1921 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1981 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2041 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2101 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2161 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2221 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2281 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     2341 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2401 actaggggtt cctgcggccg cacgcgttgt aagagtaata atcgcttata atcaggcttc
     2461 cagcctccat taaagtagac cttgacaacc atttgcttgg tctcactccc ttttctcacc
     2521 cgtttctctt tcaggcttgg tccccctcga cccccccaaa taactaggtc ctgtgggtca
     2581 ggatagttga tgctgagaac atcagctgag tcccaccata ctcttctgct gtgttatgtc
     2641 acctatttgc aaggctggat ttcaaaacta tctggtttta agaaggactg ttattttgga
     2701 tctctaacac ctctaagtta agctggagtg aaaaagaaac ataagcagtg agtgctctga
     2761 gaactgtcct tggcatatga agacagcttc atattctgta aagtggcaaa ggtgtcaact
     2821 atggactgcc agaatgacag gcctttggac ctttttctgc ggtctgaact gcaaaagcct
     2881 aacctttgtt ctgaagtcat gtggaatatt taccctttgg tctgatagaa cgtcagatca
     2941 ccagggtctg gatatgacac attcaggtca agtgtacttc aggcactttg gtatgcttca
     3001 gtcaagatgc caatggcaag tatgaaaggt gagagaccga agggaaggaa gaggccagag
     3061 cataatcaca ggcaatacct tctagtctat ttttcggagc tcgggctggg cataaaagtc
     3121 agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg
     3181 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3241 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3301 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     3361 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     3421 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3481 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3541 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3601 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     3661 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     3721 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3781 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     3841 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3901 gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct
     3961 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4021 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4081 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     4141 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga
     4201 gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4261 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4321 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4381 gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc
     4441 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4501 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4561 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4621 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4681 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     4741 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4801 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4861 aaaacttgat ttgggtgatg gttcacgtag tgggc
//
