LOCUS       sequence-432555-        5316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2400m 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 5316)
  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 5316)
  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: AiP1729 - pAAV-AiE2400m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..153
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   24..153
                     /label=AAV2 ITR
     polyA_signal    271..382
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     569..589
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(682..1392,1393..1395,1396..1398))
                     /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(724..745)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1073..1092
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1332..1353
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1395..1404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2602..2731
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2793..3381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2873..2892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3552..4343,4344..4412))
                     /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     4175..4194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4413..4517)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4585..4603
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4641..4663)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4763..4782
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4799..5254)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4936..4957)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5148..5167
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
       61 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
      121 agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgagatc
      181 tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
      241 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
      301 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
      361 ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt cagtgcccaa
      421 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaag
      481 aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
      541 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
      601 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc gaattcgcgg
      661 ccgcggcgcg ccgtcgactt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
      721 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
      781 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
      841 gttctgctgg tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt
      901 gaagttggcc ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt
      961 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     1021 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1081 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     1141 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta
     1201 gcccagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag
     1261 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     1321 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     1381 ctcgcccttg ctcaccatgg tggcgaccgg tagcgctagc ttcgaaagat ctggatccca
     1441 gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc cgagctccgc
     1501 ctctctgggg agatcaatag ttaaatatta ttgctttaag tagttcaaac aatcatctca
     1561 ggtcttagac agtaattttc agacttcttg aatgtcctcg catttattct ggtattaagc
     1621 taatcaggtg gttgcaatta ctggaaagag tctgtataga actaattatt cagttggaat
     1681 aattgtgggc aactaggcaa gcagccatca tgtgattccc ccactgaagg agccttcatg
     1741 gctgaacggc tgagctgtgc acatcattcc tgagtgtgtt taaactcact gaccatactg
     1801 cactctgcca ggctctgcgt cctcttcata gctgggtgtc ctggattttc cagataattg
     1861 ctccactcct aaattttctg ttagtatata gttgagtttg ttgtaggaaa tgttagaaca
     1921 tctctcgttt atttttgagc ccgactgaaa gtgtgaaaaa ggattctatt taaaggctga
     1981 agctgtcaat gcgcttgtga gagtgatggg tatcatatta ttcatggtag gacttagtat
     2041 cttataaaat ggtgtcacag atattcatcc atgaacagtt aagttagata ttcttaatta
     2101 tggtagaatt tgaaatcatt taaatgatcc ggaggagcca atgagtgcct acacaatttg
     2161 aagggctgtc tgtgtggtgg agatggaaga gtctcagatc cttagtctgc caacagcaga
     2221 agttacaatg tggcggtctg tgtcttggaa tggaaacatc agcattctcc cattttcaaa
     2281 agtattgagt ggcttctttc tcttttgctt tccattctca tctatagagc cgctgcagct
     2341 tcgcctgcct gggactttag attatcgtac tgctgacagg aggatttccg ggaaagagca
     2401 gggactttgg cactgagttg aaacttcaaa gcctcaactt ttcccctctc agtgtatttg
     2461 ttgggtgacc agtcttagcc tccttgtttg atcatgaata ctgcaaggcc tttatgataa
     2521 tcacaagcga tgtatgtgtg aagcaacatt ccctgcctgt gtacatgtgc cgattattac
     2581 tcttacaacg cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2641 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca
     2701 gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc
     2761 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2821 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2881 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2941 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3001 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3061 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3121 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3181 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3241 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3301 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3361 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3421 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3481 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3541 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3601 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3661 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3721 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3781 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3841 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3901 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3961 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4021 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4081 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4141 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4201 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4261 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4321 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4381 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4441 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4501 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4561 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     4621 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4681 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4741 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa
     4801 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4861 tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata
     4921 gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4981 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa
     5041 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5101 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5161 aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5221 cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt
     5281 gtgaaatacc gcacagatgc gtaaggagaa aatacc
//
