LOCUS       sequence-433041-        5104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2402m 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 5104)
  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 5104)
  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: AiP1706 - pAAV-AiE2402m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    223..334
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     521..541
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(634..1344,1345..1347,1348..1350))
                     /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(676..697)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1025..1044
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1284..1305
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1347..1356
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2342..2471
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2533..3121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2613..2632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3292..4083,4084..4152))
                     /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     3915..3934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4153..4257)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4325..4343
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4381..4403)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4503..4522
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4539..4994)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4676..4697)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4888..4907
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5069..5104,1..105)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(5080..5104,1..105)
                     /label=AAV2 ITR
ORIGIN
        1 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
       61 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc
      121 gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
      181 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
      241 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
      301 caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag ccacggaatt
      361 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
      421 cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag ccatacggga
      481 agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac
      541 atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt gattatgata
      601 tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca tgccgagagt
      661 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
      721 tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc
      781 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt
      841 gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac
      901 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
      961 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     1021 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     1081 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca
     1141 ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     1201 ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     1261 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     1321 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag
     1381 atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt ttatgcccag
     1441 cccgagctcc gaaatgcaca aggatcttca tagaaccata atttttaaaa agcaaacact
     1501 tcccccaaag gagcatgtaa gacaacttga aggtattaaa atgggattgt ggttttatta
     1561 gttagagaac atcatgcaaa ttatattatc taggcattat aatgcaaact ttaaaaggaa
     1621 atgctcatta tgtaatgtta agtgaaaaaa atcaggccat aaaatgaaat gtgcattatg
     1681 catgcatatg atgaaaagac aagaaataga taagaatatg cagaaataca aatagttggc
     1741 tgtggtgtgg gcttctggct tgttttctgt tttaaagcca ttacatcatc tttccaatga
     1801 caaactacat tttaagttat tgaaaacaaa atctcgttag ccttgtcctt gcataaattc
     1861 ccagagaaca cattattctt gttcccttct acctctgcta cttgtctttg atttggacct
     1921 gctccacctc ctcttacttt tcagtcctga actagatgga aaaaagaaat agaccctcag
     1981 gaaacaagga agcaaaacca cccaactgtg tggaagccgt gcattcaagc tgcaaacgag
     2041 tggagtggct tcaatacctt aacttatccc cattgttgca aagggagacc cagattcttg
     2101 aatgaccaca agtggcctcg gtttaacaag ccttgccaat gaggcacaat gttcccgtcc
     2161 tcagaaatag agcccggcag agcttccaaa gacccataaa agtctgcact tttggaagga
     2221 tgtgacaacc ctggcaacaa gcacagattc gtcattctct atattaagag aacgtttggt
     2281 tgcatcaggc ttttgagctg ggccaacact cgattattac tcttacaacg cgtgcggccg
     2341 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2401 gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2461 ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2521 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2581 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2641 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2701 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2761 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2821 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2881 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2941 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3001 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3061 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3121 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3181 agggattttg gtcatgagat taacaaaaag gatcttcacc tagatccttt taaattaaaa
     3241 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3301 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3361 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3421 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3481 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3541 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3601 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3661 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3721 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3781 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3841 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3901 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3961 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4021 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4081 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4141 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4201 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4261 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4321 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4381 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4441 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4501 tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt
     4561 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat
     4621 cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt
     4681 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4741 ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag
     4801 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4861 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc
     4921 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4981 gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc
     5041 gtaaggagaa aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact
     5101 gagg
//
