LOCUS       sequence-466286-        4746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2349m 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 4746)
  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 4746)
  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: AiP1670 - pAAV-AiE2349m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..56)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     156..178
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(216..234)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        302..406
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(407..475,476..1267)
                     /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(625..644)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1438..2026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1927..1946
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2088..2217
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2845..2854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2851..2853,2854..2856,2857..3567)
                     /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(2896..2917)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3157..3176)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3504..3525
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3660..3680)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3867..3978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4096..4236
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4096..4225
                     /label=AAV2 ITR
     rep_origin      join(4311..4746,1..20)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4398..4417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4608..4629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
       61 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
      121 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
      181 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
      241 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
      301 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      361 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      421 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      481 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      541 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      601 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      661 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      721 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      781 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
      841 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
      901 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
      961 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1021 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1081 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1141 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1201 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1261 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1321 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1381 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1441 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1501 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1561 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1621 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1681 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1741 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1801 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     1861 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1921 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     1981 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2041 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
     2101 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
     2161 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     2221 gccgcacgcg ttgtaagagt aataatcgaa gccaaggcag gtgaaagact ggatttaaga
     2281 tccagaacca ctagaagaga acaggaaaag cacttcaaga cgtccctctt ggaactgatt
     2341 ttctgtgtca cacccaaaac aacccacagg aaaccgacac aaaaagtgac aaatgcctat
     2401 gataacaaat ccaaatcagg aagctactgc acagcaaagg gacagatgag agagtgaaga
     2461 gacaccctgc atagcagaag actgtctgcc aaccacttgt cacatccaga atatatgcag
     2521 actaccttta ctacagtgga gagcatgtgt ggagagcatg agtgtcaaat tacatctgaa
     2581 cggtagatac tccataccta caattacctc taatttaaac ccccaaactc ctgacaatga
     2641 cataactctg atcaggcatt cacactgacc cattaaacat gtttaggggc tggagtcaca
     2701 ctgaaccaag atgatggcta cctcgcaatt catgatgaga gtgtgttcac ggagctcggg
     2761 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
     2821 ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     2881 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2941 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     3001 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     3061 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3121 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3181 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3241 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3301 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     3361 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3421 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     3481 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3541 actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     3601 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3661 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3721 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     3781 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3841 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     3901 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3961 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4021 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     4081 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4141 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     4201 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     4261 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4321 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4381 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4441 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4501 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4561 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4621 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     4681 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4741 taacaa
//
