LOCUS       sequence-432585-        5262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2437m 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 5262)
  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 5262)
  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: AiP1916 - pAAV-AiE2437m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(108..212)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     280..298
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(336..358)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     458..477
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(494..949)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(631..652)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     843..862
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1024..1164
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1035..1164
                     /label=AAV2 ITR
     polyA_signal    1282..1393
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1580..1600
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1693..2403,2404..2406,2407..2409))
                     /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(1735..1756)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2084..2103
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2343..2364
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2406..2415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3559..3688
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3750..4338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3830..3849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4509..5262,1..38,39..107))
                     /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     5132..5151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
       61 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
      121 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
      181 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
      241 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
      301 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      361 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      421 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      481 agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
      541 taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa
      601 gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
      661 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
      721 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
      781 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
      841 gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg
      901 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc
      961 tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     1021 gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
     1081 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     1141 ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg
     1201 ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa
     1261 tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga
     1321 gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa
     1381 ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca acagccgagc
     1441 ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca
     1501 ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg
     1561 cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa
     1621 tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg gccgcggcgc
     1681 gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     1741 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta
     1801 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     1861 gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc
     1921 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta
     1981 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     2041 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     2101 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     2161 gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt
     2221 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt
     2281 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     2341 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     2401 gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga tctggatccc agaagcaaat
     2461 gtaagcaata gatggctctg ccctgacttt tatgcccagc ccgagctccg atccccctga
     2521 gaaatgagta agagaacagg attgcaaggc agtcggtggt gcaaacctta acactcagtt
     2581 tctctatcta gtcactggtt agtaacccca tgccctcagt ggataagggg gtgggggggg
     2641 gaatgttggt tttatctgta gataaaacag gctagccatt gatgaaggtt tcttcccttt
     2701 ctcctccaat agctgtgaga agccacaatc tagaaatagc tacaatgccc ataaatgctg
     2761 tcaccagagg aagggtcctg cctgtgacac aaaagacagg tcctgcttgg ttgctggcat
     2821 cttgggggca gagctgaatt cctgggctga tgagtttggc gagggtgggc agccaccctc
     2881 ctggctccct cttcttcctc tcaccatctc ttccaaccac gtgtctgtct ttccaacccc
     2941 ctcagaacgt gtgtttgtag ccagcttctc agtggcctag ggaggatttt taaaaatagc
     3001 gtcattgatt tcatgttttg gaataagaaa agcaatagac gcgttctgtt tagaaaaaaa
     3061 aataattaga tggtacagaa aagaaaaata aaaaagagag gggaattggc ctaaatcttc
     3121 tccacagaaa ctttcatcat ccctatttgg tatggtgccc cacccccaat ccgagtcctg
     3181 ttctctgtgg ccgtcctgtg ggcttccttg tccagatggg atgggggcct gactctatgg
     3241 cagatggatc agagctggcc agcaggtgtg aatccagcat agtgtggtgc agtgtggaca
     3301 gacccacagg gccaccctgg ttcttcagcc tagtcatcgc tgcatagtca cttccccaca
     3361 acctgcctta tcagaatact ctccatcgat acctttccgg ttcacctgga gtcttacgaa
     3421 atgaatgcca cacagctaac taaggcctgc gtcagctttg ttcagagggg caagacgact
     3481 ctgtccaagg catagcaaag ttgaagtgag gaagccagcc aggtctctga tggtattaga
     3541 cttgaagtgt gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3601 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg
     3661 agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg
     3721 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3781 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3841 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3901 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3961 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4021 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4081 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4141 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4201 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4261 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4321 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4381 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4441 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4501 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4561 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4621 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4681 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4741 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4801 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4861 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4921 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4981 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5041 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5101 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5161 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5221 ttgagatcca gttcgatgta acccactcgt gcacccaact ga
//
