LOCUS       sequence-432541-        4948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2168m 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 4948)
  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 4948)
  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: AiP1439 - pAAV-AiE2168m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(66..170)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     238..256
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(294..316)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     416..435
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(452..907)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(589..610)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     801..820
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   982..1122
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   993..1122
                     /label=AAV2 ITR
     polyA_signal    1240..1351
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1538..1558
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1651..2361,2362..2364,2365..2367))
                     /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(1693..1714)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2042..2061
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2301..2322
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2364..2373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3203..3332
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3394..3982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3474..3493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4153..4944,4945..4948,1..65))
                     /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     4776..4795
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
       61 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
      121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      181 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
      241 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
      301 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
      361 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
      421 tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat tttgttaaaa
      481 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga aatcggcaaa
      541 atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc agtttggaac
      601 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
      661 ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc gaggtgccgt
      721 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
      781 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca
      841 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
      901 ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga
      961 gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg
     1021 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     1081 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca
     1141 cgtgagatct catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc
     1201 ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat
     1261 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
     1321 acaacagatg gctggcaact agaaggcaca gtcgaagatc tctcgagcca cggaattgtc
     1381 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
     1441 cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc
     1501 aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata
     1561 gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatgatatcg
     1621 aattcgcggc cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc cgagagtgat
     1681 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     1741 gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     1801 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg
     1861 gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt
     1921 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     1981 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     2041 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     2101 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg
     2161 cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     2221 gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     2281 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     2341 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagct tcgaaagatc
     2401 tggatcccag aagcaaatgt aagcaataga tggctctgcc ctgactttta tgcccagccc
     2461 gagctccgcc ggcgaccagc aagggaccaa ggaaaggcaa atgggggcga ggccacaaga
     2521 agtgttttag tgctgggtca gctggtatga gggcctggct caagtctgga accagaacca
     2581 gtctgggcta ctgtaagggc aggtcccttc cgttcttggc ctccttgtcc tagatgtggt
     2641 agttgagtgt gatgaccccc cccccatgtc ttccggtaga gcctgtcatg gtaaccgcct
     2701 gctctgggga gccgagtgag tcatgagtga cgggatggtt actcaccgac cacctggctt
     2761 gggggtgcag agggtgctca ccactgtcct cccaggctca ggcagacctc ttcctggttc
     2821 ctgctgtggc tttgctggtg gtgctgacca cagtcctaaa agccctgtgt gtatgaccct
     2881 tggtgagatg ggcgctgaat cagtctgcta ctggggcatc acagtcgggg gtgggggaca
     2941 gtgtgggcca caggacttcg acatgaacag gtcacctgga gggtggtgtc cgtctggtcc
     3001 tcagactcct cataagtcag tcgtagggga aagaagactc aggtcaggcc caggcgtctc
     3061 tgggtttgaa agcatgtgac acccactagg acaactggca caccagggtt agagccagaa
     3121 caccggagaa cagcttaagc cccaggaaac cttccagctg gagcccgccg gcgattatta
     3181 ctcttacaac gcgtgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3241 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc
     3301 agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc
     3361 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3901 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3961 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4021 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4081 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4141 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4201 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4261 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4321 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4381 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4441 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4501 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4561 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4621 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4681 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4741 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4801 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4861 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4921 tactttcacc agcgtttctg ggtgagca
//
