LOCUS       sequence-466305-        4555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2501m 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 4555)
  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 4555)
  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: AiP1781 - pAAV-AiE2501m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(203..791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(283..302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(962..1753,1754..1822))
                     /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     1585..1604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1823..1927)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1995..2013
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2051..2073)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2173..2192
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2209..2664)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2346..2367)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2558..2577
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2739..2879
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2750..2879
                     /label=AAV2 ITR
     polyA_signal    2997..3108
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3295..3315
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3408..4118,4119..4121,4122..4124))
                     /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(3450..3471)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3799..3818
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4058..4079
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4121..4130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
       61 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc
      121 gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      181 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      241 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      301 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      361 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      421 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      481 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      541 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      601 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
      661 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      721 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      781 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      901 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      961 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1021 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1081 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     1141 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1201 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1261 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1321 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1381 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1441 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1501 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1561 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1621 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1681 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1741 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1801 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1861 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1921 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     1981 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     2041 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2101 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     2161 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg
     2221 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2281 agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg
     2341 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2401 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt
     2461 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2521 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2581 gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2641 ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc
     2701 gcacagatgc gtaaggagaa aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg
     2761 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc
     2821 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
     2881 cggccgctcg gtccgcacgt gagatctcat gcctgctatt gtcttcccaa tcctccccct
     2941 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
     3001 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     3061 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaagatctct
     3121 cgagccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag
     3181 gcggcgatga gttccgccgt ggcaagaact aaccaggatt tatacaagga ggagaaaatg
     3241 aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc
     3301 gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga
     3361 ggttgattat gatatcgaat tcgcggccgc ggcgcgccgt cgacttactt gtacagctcg
     3421 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     3481 ttctcgttgg ggtctttgct cagcttggac tggtagctca ggtagtggtt gtcgggcagc
     3541 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg
     3601 ccgtcctcga tgttgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttgtcg
     3661 gcggtgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     3721 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     3781 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     3841 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     3901 tagcgggcga agcactgcac gccgtagccc agggtggtca cgagggtggg ccagggcacg
     3961 ggcagcttgc cggtggtgca gatcagcttc agggtcagct tgccgtaggt ggcatcgccc
     4021 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     4081 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtagc
     4141 gctagcttcg aaagatctgg atcccagaag caaatgtaag caatagatgg ctctgccctg
     4201 acttttatgc ccagcccgag ctccgtttac ccattacatc ttttacccat atgattagaa
     4261 aacaaaatcc tgttaaatcc cagagctgtc tgcctcaggg atccttttgt tttggatgaa
     4321 taggtgccct cagcccattt aatttcactt tggacgcaga gagcttaata taaggaccag
     4381 cctttgaact tggagaaagc tgagagacca cagctacacc tgcctatcag taaaataatc
     4441 tgaaagccat tttgaggggg accatctgaa caatcctatt aggagtgatc catttagaag
     4501 tgggatcaaa gacttaagat atttatggca gatatcgatt attactctta caacg
//
