LOCUS       sequence-466253-        5337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2163m 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 5337)
  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 5337)
  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: AiP1392 - pAAV-AiE2163m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      563..572
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(569..571,572..574,575..1285)
                     /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(614..635)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(875..894)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1222..1243
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1378..1398)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1585..1696
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1814..1954
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1814..1943
                     /label=AAV2 ITR
     rep_origin      2029..2484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2116..2135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2326..2347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2501..2520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2620..2642
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2680..2698)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2766..2870
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2871..2939,2940..3731)
                     /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(3089..3108)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3902..4490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4391..4410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4552..4681
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aaatggattt atactgtctt tgaacactat tgagaccaaa ttgctatttt atattttgtg
       61 tcattttgtt cttttatttt attgctttct ttgaggttaa atagatattt gcgtgtgtta
      121 gtttaatgcc tttgtctttt cttgtaattt attttaaggc tttttttttt tattggttgc
      181 cttgaagatt ataactacta tagtagtaca tgtgtgtgtg ctcatgtgtg tgtgtgcatg
      241 ggtgtatgtg gcatttgcgt acctgaatgc atgtgtacag ttatacatgt ctgtgcatgc
      301 acacagagaa gccagacact agtgtcaggt gtcctgccct gccactttcc cacattttga
      361 gactgcattt acagctcagc aatacaatta tcccggctgg ccagggagcc ttcgagggtt
      421 tcctgtctct gccttccagt atcgagcaca gggttacaag ccgccggcgg agctcgggct
      481 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
      541 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
      601 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      661 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
      721 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
      781 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      841 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
      901 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
      961 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1021 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1081 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1141 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1201 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1261 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     1321 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1381 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1441 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     1501 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     1561 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1621 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1681 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1741 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     1801 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     1861 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     1921 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     1981 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2101 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2221 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2341 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     2401 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2461 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     2521 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2581 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     2641 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     2701 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2761 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2821 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2881 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2941 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3001 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3061 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3121 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3181 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3241 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3301 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3361 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3421 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3481 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3541 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3601 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3661 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3721 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3781 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3841 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3901 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3961 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4021 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4081 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4141 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4201 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4261 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4321 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4381 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4441 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4501 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
     4561 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
     4621 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     4681 tgcggccgca cgcgttgtaa gagtaataat cgccggcgcc ttgctcctgg agttggggga
     4741 tgagattggt cccagaaaaa ctttgaacac cggtgacacc agggagggaa gaagagcctt
     4801 aacaacagca acatcacaag acaagaaaca aatcttagga aacatggttg gattgtcctt
     4861 ttcagagctc tgaaaattca tgaacctttt tcagtaatca gaagagcaat ctgctaaaga
     4921 aaagagggaa aggacggtgt gtgtgtgtgt gtgtgtgtgt gtgtgtaacc ggagctctct
     4981 agcactagct ggccttaccc ccacaccctc cccagcccag tagtgtccca gtgaaagcaa
     5041 caagcagcgc tctgagtagc aggagaactc ctccacattc atgtctgcct tcctaggccg
     5101 gccgcaggcc ccctggaagg accccactta caaggctgtc tttatctgcc tgactcatag
     5161 ttcacacagt gggaaagcct ttgcctggag gtgtgtttag tttctgtttg cgacacttct
     5221 tctctcctga gttaaaagac aactgcggga agcaataatt aggaatatgt gttgataggc
     5281 atgcaatata caaaggtata atttgtgtga caataacggc ccaaaggaca gcggaga
//
