LOCUS       sequence-466288-        4794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2358m 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 4794)
  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 4794)
  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: AiP1676 - pAAV-AiE2358m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   212..352
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   223..352
                     /label=AAV2 ITR
     polyA_signal    470..581
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     768..788
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(881..1591,1592..1594,1595..1597))
                     /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(923..944)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1272..1291
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1531..1552
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1594..1603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2279..2408
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2470..3058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2550..2569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3229..4020,4021..4089))
                     /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     3852..3871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4090..4194)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4262..4280
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4318..4340)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4440..4459
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4476..4794,1..137))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4613..4634)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa
       61 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
      121 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
      181 tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc
      241 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
      301 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
      361 tcggtccgca cgtgagatct catgcctgct attgtcttcc caatcctccc ccttgctgtc
      421 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
      481 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
      541 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaagatc tctcgagcca
      601 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      661 tgagttccgc cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca
      721 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
      781 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
      841 tatgatatcg aattcgcggc cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc
      901 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
      961 tggggtcttt gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg
     1021 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct
     1081 cgatgttgtg gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga
     1141 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     1201 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     1261 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     1321 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg
     1381 cgaagcactg cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct
     1441 tgccggtggt gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     1501 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     1561 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagct
     1621 tcgaaagatc tggatcccag aagcaaatgt aagcaataga tggctctgcc ctgactttta
     1681 tgcccagccc gagctccgct ttgagttgac agaaaaggta tggctaaccc taacattgac
     1741 acctgccaga gctttctgca tatcaaaaag atggagacat agaattagag ttatagcttt
     1801 gagagtatta cctgtgttta gaaatcccag ttaactctag taccataaga taagtacttc
     1861 atgtaactta tttgtgaaga aataaaatgg aaactcactt actcatgaac accaatgcct
     1921 ttcaacaagc agaatctgtt ttgctggaag tctaaattga gaaaaattta acactttttt
     1981 ttttcagcaa ggacttgaaa gacgagatag ctgttttggt ggaaaagaag agacacaaag
     2041 tttccacagc tgagaaactg gaagactgta tggattttgc aactgatttg attttcgctg
     2101 aggtattggc ctgaatgaac cacaattcct cacctgatat gtattatggc agtgtaccat
     2161 tatttagaaa gtccttttta gaactgtggc atgatttctc tttaaaaagt tcctctagca
     2221 gccttatgat agcccatcaa aacagcccga ttattactct tacaacgcgt gcggccgcag
     2281 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     2341 gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     2401 cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3301 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3361 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3421 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3481 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3541 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3601 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3661 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3721 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3781 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3841 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3901 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3961 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4021 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4081 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4201 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4261 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4321 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4381 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4441 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt
     4501 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg
     4561 caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg
     4621 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4681 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
     4741 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacg
//
