LOCUS       sequence-432569-        4756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2530m 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 4756)
  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 4756)
  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: AiP1851 - pAAV-AiE2530m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(97..201)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     269..287
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(325..347)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     447..466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(483..938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(620..641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     832..851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1013..1153
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1024..1153
                     /label=AAV2 ITR
     polyA_signal    1271..1382
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1569..1589
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1682..2392,2393..2395,2396..2398))
                     /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(1724..1745)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2073..2092
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2332..2353
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2395..2404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3042..3171
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3233..3821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3313..3332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3992..4756,1..27,28..96))
                     /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     4615..4634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
       61 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
      121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
      181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
      241 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
      301 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      361 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      421 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      481 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      541 attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      601 gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      661 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      721 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      781 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      841 agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      901 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
      961 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg
     1021 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     1081 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     1141 actaggggtt cctgcggccg ctcggtccgc acgtgagatc tcatgcctgc tattgtcttc
     1201 ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact
     1261 cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc
     1321 cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac
     1381 agtcgaagat ctctcgagcc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     1441 agcgggcaag gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca
     1501 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     1561 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     1621 ttttgtaatc cagaggttga ttatgatatc gaattcgcgg ccgcggcgcg ccgtcgactt
     1681 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     1741 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactggtag ctcaggtagt
     1801 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     1861 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt
     1921 tcttctgctt gtcggcggtg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     1981 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     2041 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     2101 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     2161 tcatgtggtc ggggtagcgg gcgaagcact gcacgccgta gcccagggtg gtcacgaggg
     2221 tgggccaggg cacgggcagc ttgccggtgg tgcagatcag cttcagggtc agcttgccgt
     2281 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     2341 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     2401 tggcgaccgg tagcgctagc ttcgaaagat ctggatccca gaagcaaatg taagcaatag
     2461 atggctctgc cctgactttt atgcccagcc cgagctccgc ccaggagcag ttcttaatat
     2521 tttgccctcc aaagcagagg gaggtgaggt tctctttgtt caacttctct cctgcttctc
     2581 tcttctcaaa tctaattgac tctagatgtt tgatctctgg tttttataga tgaatgcttt
     2641 taaacttttc ataaaaggtg ccgcataaac ccttcagaaa catttcattc gacatcgcgc
     2701 tttcaagatt tacctagtat gaacatgtct gcatcgttct cattgctgga catttaaatt
     2761 cattacactc cctgcggaca cacagcagtc tgccagattc tacatctaga acacgcctgc
     2821 caagtcctgg cagaagtgct gccaactggt ctccaaggag actgcaccaa ttttcactcc
     2881 cagcagtagt gagttcctgg ggttaagctt gctcatcaat agcccacgtt taaaattttc
     2941 tgtccatctg aattggacaa aaatggtatc ttaacatgca ttgctctcag ccagcgcttc
     3001 acctttctct cgattattac tcttacaacg cgtgcggccg caggaacccc tagtgatgga
     3061 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3121 ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag
     3181 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3241 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3301 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3361 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3421 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3481 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3541 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3601 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3661 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3721 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3781 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3841 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3901 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3961 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4021 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4081 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4141 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4201 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4261 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4321 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4381 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4441 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4501 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4561 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4621 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4681 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4741 actgatcttc agcatc
//
