LOCUS       sequence-462001-        5221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2439m 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 5221)
  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 5221)
  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: AiP1768 - pAAV-AiE2439m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1233..1242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1239..1241,1242..1244,1245..1955)
                     /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(1284..1305)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1545..1564)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1892..1913
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2048..2068)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2255..2366
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2484..2624
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2484..2613
                     /label=AAV2 ITR
     rep_origin      2699..3154
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2786..2805)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2996..3017
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3171..3190)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3290..3312
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3350..3368)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3436..3540
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3541..3609,3610..4401)
                     /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(3759..3778)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4572..5160
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5061..5080
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaag agtaataatc gattaactta tgtgtttctt
      181 tgaatttcaa gttgaaatgg ctcctgtgtt gatgtcattc aaccatggct gacagaaaat
      241 agcaatattc actttcatca aagttatatg tgatttagga aaataattaa aagccagtgg
      301 caaagataga aactctactc ataagcctat tcagatttat atctgctcca acatttgcag
      361 tcatataata gatgtttctt ttagagttat ttcagttcat ttcaataaga aacagtagct
      421 aagggaaaaa ttcatggcat tacaattgat attaaaagaa ccaatgagct ttgccactaa
      481 tctgagcact tttatttact tcagagaatg ataccgatta ttaagcacat actttcttcc
      541 atttttgttg ttcctgattg gtgccagggt tttcttggct tgttgcggga taggctgaat
      601 agaccaactt tcctttgcat ctgcatattc gataggtcta gtgattctga atccaagtca
      661 cttgaataat gatggaacca ttttgttcct gatgaataca cactaatata atcagatgct
      721 gtcaaaagaa agaaagaaaa ctcatgtgaa ccagaatggt gcatacaatc atccattgct
      781 accaaaccga agggatgctt aggcgtacag ggccaagatc ctttggtaaa tgaaccacat
      841 agctgagtaa taataatgtg taaaatatta actacttgaa aaatactcta ttattgaata
      901 aagcactctc cactcactca aactagacca gaagaagtct gggaagcagt atgattagag
      961 agagtgggtg gaaaagaatc atttctcagc tgggattcaa atggtctctt gggaaattta
     1021 tagtttgaaa ggaatttagg atctcgaaaa ctaaaaatgt taaagaccaa gtacatgaca
     1081 gtattcacat taaaactcag taggttagga ggaacacatg ttatcaggag agcatgtcgg
     1141 agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg
     1201 atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1261 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1321 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
     1381 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
     1441 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1501 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1561 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1621 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1681 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     1741 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1801 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     1861 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1921 ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc
     1981 gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2041 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2101 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg
     2161 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2221 gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct
     2281 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2341 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2401 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc
     2461 tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     2521 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     2581 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     2641 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     2701 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2761 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2821 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2881 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     2941 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3001 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     3061 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3121 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     3181 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3241 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3301 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3361 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3421 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3481 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3541 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3601 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3661 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3721 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3781 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3841 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3901 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3961 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4021 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4081 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4141 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4201 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4261 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4321 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4381 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4441 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4501 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4561 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4621 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4681 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4741 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4801 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4861 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4921 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4981 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5041 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5101 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5161 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5221 t
//
