LOCUS       sequence-432530-        4658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2113m 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 4658)
  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 4658)
  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: AiP1331 - pAAV-AiE2113m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(337..1128,1129..1197))
                     /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     960..979
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1198..1302)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1370..1388
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1426..1448)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1548..1567
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1584..2039)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1721..1742)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1933..1952
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2114..2254
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2125..2254
                     /label=AAV2 ITR
     polyA_signal    2372..2483
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2670..2690
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2783..3493,3494..3496,3497..3499))
                     /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(2825..2846)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3174..3193
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3433..3454
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3496..3505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4045..4174
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4236..4658,1..166))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4316..4335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
       61 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      121 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      181 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      241 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
      301 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
      361 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
      421 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
      481 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
      541 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
      601 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
      661 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
      721 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
      781 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
      841 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
      901 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
      961 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1021 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     1081 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     1141 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     1201 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1261 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     1321 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     1381 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     1441 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     1501 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     1561 gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt
     1621 aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta
     1681 taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc
     1741 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1801 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1861 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1921 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc
     1981 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
     2041 ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2101 cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca
     2161 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     2221 gagggagtgg ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat
     2281 ctcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     2341 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     2401 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     2461 tggctggcaa ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca
     2521 acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa
     2581 gaactaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     2641 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     2701 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg
     2761 gccgcggcgc gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg
     2821 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct
     2881 tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     2941 tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct
     3001 tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt
     3061 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     3121 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     3181 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     3241 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt
     3301 agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca
     3361 gcttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     3421 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     3481 cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga tctggatccc
     3541 agaagcaaat gtaagcaata gatggctctg ccctgacttt tatgcccagc ccgagctcaa
     3601 actcagtgaa atagtataat tactatagtc tatttaaaaa gattattaat aaagggaaat
     3661 atcctaaaag attctactac ttgtttaaaa agatgaagtc tgagtaccac taattttcag
     3721 aatttattct taggatctag gtactgttct gcctctaaat ttaatcttca cctcaatgct
     3781 tctctacatc acagttaaat aactcctaga acagttccgc tctcccttgc gatctgtgtg
     3841 cttagggtgc tgccaagctg aattcaggga gcagctgtgt tttaaactct gagaggttat
     3901 acctttagat agatgagatg ggatctcttg gagatatggg agtctgcaca ctgcaggaga
     3961 tgtgtctcca ctggcttcct cacacaccct gcctctgcac cggacagccc cagtcccaag
     4021 ttcatgctac acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4081 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc
     4141 tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag
     4201 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4261 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4321 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4381 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4441 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4501 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4561 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4621 gtaggcggtg ctacagagtt cttgaagtgg tggcctaa
//
