LOCUS       sequence-466276-        4758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2280m 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 4758)
  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 4758)
  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: AiP1613 - pAAV-AiE2280m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(141..160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     260..282
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(320..338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        406..510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(511..579,580..1371)
                     /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(729..748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1542..2130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2031..2050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2192..2321
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2961..2970
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2967..2969,2970..2972,2973..3683)
                     /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(3012..3033)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3273..3292)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3620..3641
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3776..3796)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3983..4094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4212..4352
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4212..4341
                     /label=AAV2 ITR
     rep_origin      join(4427..4758,1..124)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4514..4533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4724..4745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
       61 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
      121 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
      181 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      241 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      301 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
      361 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      481 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      541 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      601 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      661 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      721 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
      781 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
      841 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
      901 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
      961 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1021 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1081 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1141 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1201 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1261 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1321 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1381 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1681 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2161 ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc
     2221 actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     2281 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca cgcgttgtaa
     2341 gagtaataat cgccggcgaa agcaccgcct atgctgtggg ttgaactgct caccgctctg
     2401 cgaaagcctg tggtttgcat tcagtgtgat tcgtcctgcc tgctgaccac tgtgctgggc
     2461 ttacacttct gacactgcca ggctacccaa cttgtggttc tgtggttgtt tatgaggtcc
     2521 aaagaagttt tcacacaacc caaattacaa atttaactgt tcccctttcc acagcccacc
     2581 gccattggct cttgccaatc acgtgactta agtgatgtca attttttttt tttaaacttt
     2641 ctgagctatg cccttcctgc cctccacctg ccctccccca ggctgtgtaa ggaaacagtt
     2701 gagcagagct tagcagagag gggctgtaag agaaaaaaaa gcacaacaaa caagcaaaca
     2761 gaaatcccct gcctcagtct tttcttgctc tctctgtcct gtgcggattt ggtggctttc
     2821 agatcctgtt gttatttgtg gtttgaggaa caactgattc gccggcggag ctcgggctgg
     2881 gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat ccagatcttt
     2941 cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3001 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3061 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
     3121 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct
     3181 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3241 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3301 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3361 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3421 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     3481 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3541 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     3601 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3661 tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc
     3721 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3781 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3841 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     3901 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3961 ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4021 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4081 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4141 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     4201 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4261 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4321 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     4381 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     4441 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4501 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4561 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4621 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4681 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4741 actggaacaa cactcaac
//
