LOCUS       sequence-433019-        5543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2169m 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 5543)
  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 5543)
  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: AiP1398 - pAAV-AiE2169m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(16..604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(96..115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(775..1566,1567..1635))
                     /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     1398..1417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1636..1740)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1808..1826
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1864..1886)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1986..2005
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2022..2477)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2159..2180)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2371..2390
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2552..2692
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2563..2692
                     /label=AAV2 ITR
     polyA_signal    2810..2921
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3108..3128
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3221..3931,3932..3934,3935..3937))
                     /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(3263..3284)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3612..3631
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3871..3892
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3934..3943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   5368..5497
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
       61 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      421 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1501 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1621 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1741 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1801 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     1861 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     1921 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     1981 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat
     2041 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga
     2101 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc
     2161 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2221 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     2281 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2341 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa
     2401 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2461 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
     2521 tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc
     2581 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     2641 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
     2701 tcggtccgca cgtgagatct catgcctgct attgtcttcc caatcctccc ccttgctgtc
     2761 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     2821 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     2881 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaagatc tctcgagcca
     2941 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
     3001 tgagttccgc cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca
     3061 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
     3121 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
     3181 tatgatatcg aattcgcggc cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc
     3241 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     3301 tggggtcttt gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg
     3361 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct
     3421 cgatgttgtg gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga
     3481 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     3541 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     3601 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     3661 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg
     3721 cgaagcactg cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct
     3781 tgccggtggt gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     3841 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     3901 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagct
     3961 tcgaaagatc tggatcccag aagcaaatgt aagcaataga tggctctgcc ctgactttta
     4021 tgcccagccc gagctccgcc ggcgcaggcc aaaaggatgt ctgggtttct atggacagct
     4081 tctcctctaa cttctggaaa gggaagtgga tggtgtgaaa agtgggaagt ggatggtgtg
     4141 aaaagcaccg tctagatgtg ggagaggagg cggacagtca gaggcggaag gtccgaggtg
     4201 gacagtggga agtggacggt catttctatc cagatctatc tggagaaggt tgtatgtttc
     4261 taaggaccgt taaccagagg gctgagagtg attgaaactt ctgaaataaa ccgtgtggac
     4321 aggattttca tgggtggttt ctcttgcttc ctatcagtgg gtctcacagg gatattccat
     4381 gaaagcatca atttgcagga gcctgagtca tgatactttc ataatttctt tcactcatct
     4441 ataatcagcc tcaagaagtt gtagaaggga gttagggaag aaagcgcctt ggcagtgatg
     4501 tgaaaaaacc ctcctggcca gcaagccact tttattcttc ctttccccct ctatatcatt
     4561 aattacacat taatattcag tgcctctcag caggcaacat cctctgctac ttgtaatcag
     4621 agagcacaca cccaggggta gggaagctat ctgtgttgct cagtagcagg atagaaggat
     4681 agaaggagcc agttctgtct ttcaagcctc aggcagggtt gagtggggag cctcctgatg
     4741 ctaacaatgt aggaagcatg tgggaactca gggtgaccac agcagcactt ttcccatggc
     4801 tcagccctca ctcacaagcc cctgctgtga tcttggtgac agtgagtagg tggatcctct
     4861 tgtgttattt ttatctgtaa cttagtaatt acttcagaat cagcagcagc acaatgcttt
     4921 gataatccca gaacaaagga agagaataaa ataatgtaag gatagatact atatgtatgt
     4981 gtctcaaaca caatatataa ttaataaaac tgtgtgtgtg tgtgcatgtg tgtgtgctag
     5041 atttaatgac cttcactgag gcaccagata tctatgatag gttcatacag aacttgcttc
     5101 tttttcagtg aataagaatt tcctttttaa gttctaaaat ctctgtgacg ttagagaaca
     5161 catttaaaga ggaaatacag cttctacaaa aatgtacact caaatttctc tatctatgac
     5221 ctccatacct acaagtccaa cttaccatga tctaaaaatt ttgtgaaaat aatttagtct
     5281 gtatggaaca tagccataca tacctcctct taccataagt ttctgagtaa cgccggcgat
     5341 tattactctt acaacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct
     5401 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg
     5461 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa
     5521 aaggccagga accgtaaaaa ggc
//
