LOCUS       sequence-432593-        4854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2586m 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 4854)
  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 4854)
  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: AiP1977 - pAAV-AiE2586m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     482..501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   643..772
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1508..1517
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1514..1516,1517..1519,1520..2230)
                     /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(1559..1580)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1820..1839)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2167..2188
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2323..2343)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2530..2641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2759..2899
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2759..2888
                     /label=AAV2 ITR
     rep_origin      2974..3429
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3061..3080)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3271..3292
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3446..3465)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3565..3587
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3625..3643)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3711..3815
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3816..3884,3885..4676)
                     /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     complement(4034..4053)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4847..4854,1..581)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
       61 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg
      661 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
      721 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
      781 agaggaagca ggaagctgca agccaagcct tagcgctttc acaacctccg tgtgcacacc
      841 ctggttcctc tgttaattgt ggcacatctg ctcctcgccc tctggatgtt tgggtccctc
      901 ggttggaatg attccaggct gtctgctcac agatgtttct cacgagtctt gtcaagtgat
      961 gcagaggtgg aaaactaaaa agaggaggca ttgtaccgtc ctccccccac cagcagtcca
     1021 ctaaaaatca gaccacccaa gcaaacacca gggctgcttt tctctgagca ttctgccaga
     1081 ttacagcaga gcgggctggg tcccgggagg gggtgtggaa gggggtgctg gcgctgaaca
     1141 gaggaaggct ttcagagcct ggacttgcca agtctcacag tctcgctttc acacattgct
     1201 tctcaatctt ggtgctgtgg ctgcctcccc aggatgcaga acatttccct cctgtgtgaa
     1261 ggatgatgat gacagcaaag tgatttgtgc aaacagtgac ctgaggttcc ctccatagac
     1321 actaaggcct gtgccacttc ccagaaaggg tcgcctccag cttgactgag atttggaaat
     1381 tctcagaatc acaaagagac cctttgagtt agcggagctc gggctgggca taaaagtcag
     1441 ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct
     1501 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1561 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1621 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
     1681 gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc
     1741 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1801 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1861 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1921 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga
     1981 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
     2041 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2101 gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg
     2161 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2221 gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct
     2281 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2341 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2401 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     2461 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga
     2521 gatcttcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2581 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2641 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2701 gggaggattg ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag
     2761 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     2821 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     2881 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2941 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3001 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     3061 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3121 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3181 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3241 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3301 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     3361 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     3421 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3481 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3541 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3601 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3661 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3721 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3781 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3841 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3901 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3961 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4021 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4081 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4141 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4201 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4261 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4321 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4381 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4441 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4501 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4561 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4621 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4681 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4741 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4801 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatc
//
