LOCUS       sequence-438967-        5707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2192m 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 5707)
  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 5707)
  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: AiP1513 - pAAV-AiE2192m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     368..387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(606..710)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     778..796
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(834..856)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     956..975
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(992..1447)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1129..1150)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1341..1360
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1522..1662
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1533..1662
                     /label=AAV2 ITR
     polyA_signal    1780..1891
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2078..2098
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2191..2901,2902..2904,2905..2907))
                     /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(2233..2254)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2582..2601
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2841..2862
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2904..2913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4502..4631
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4693..5281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4773..4792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5452..5707,1..536,537..605))
                     /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"
ORIGIN
        1 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
       61 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
      121 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
      181 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
      241 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
      301 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
      361 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
      421 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
      481 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
      541 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
      601 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
      661 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      721 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
      781 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
      841 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
      901 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
      961 tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat tttgttaaaa
     1021 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga aatcggcaaa
     1081 atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc agtttggaac
     1141 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     1201 ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc gaggtgccgt
     1261 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     1321 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca
     1381 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     1441 ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga
     1501 gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg
     1561 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     1621 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca
     1681 cgtgagatct catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc
     1741 ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat
     1801 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
     1861 acaacagatg gctggcaact agaaggcaca gtcgaagatc tctcgagcca cggaattgtc
     1921 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
     1981 cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc
     2041 aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata
     2101 gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatgatatcg
     2161 aattcgcggc cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc cgagagtgat
     2221 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     2281 gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     2341 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg
     2401 gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt
     2461 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     2521 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     2581 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     2641 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg
     2701 cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     2761 gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     2821 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     2881 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagct tcgaaagatc
     2941 tggatcccag aagcaaatgt aagcaataga tggctctgcc ctgactttta tgcccagccc
     3001 gagctccgcc ggcgcacccc tgtcaggcag ctcaaaatca cccctaacct ccagctcgag
     3061 ggacctaatg tctctcacct gtctgcactc acaggaacaa atgtgggcat gggcaagtct
     3121 taggattcca ctggggtttt ggcttttcta cttagtgagg tatccttgta tcattttcaa
     3181 atacaggcag caaagaaaaa gagctacaca gctgtaatgt catgaggctg gggatgaagc
     3241 ctcccagaca tgggatgggt ctgcttcctt cgtgcctgca gccctgggat tacaggagag
     3301 gaggccactt acaaccataa actctttgaa tcttaaaagc aattaggaag caatttgaca
     3361 aatggaaatg ggtctattta cagtttggcc aaacctggaa gaaacaggct cctggtttct
     3421 ctctttctgt aacaggctta cagttttata gacatgatga gaaaaaagcc aagaatagat
     3481 tcatgccgtt aagtttccag actgtgtgta gtttctgcct aggtggccag ttccttctct
     3541 gtctggcacc aaggcttcat tcagcttctc ccactaccaa ctgtgaaaaa ttgaattctt
     3601 tgggatccgt tgttcagtag cttgagaagg gaacagggaa gcatagcgtc tcttgagtat
     3661 atatcccctt cttcctcatt gcacagaatg catgcacact tttacatgac ttattgagca
     3721 tgtgtgccaa ggcatacctg tggaagtcag agaacacttt agaggactgg gtcttcttcc
     3781 ccttaaacca catgggttct ggggatcaac attaagtcat caggcttggg atacagttgg
     3841 ctcctgaact cctcttggac actttccttg gagcaggcct ctctgagatc ctcaggcaca
     3901 gccatggtaa ccttggttac ctttggttgc aagagaattg ggattgatgg gccaaagctg
     3961 ttctagggtg acaatcccat cacagacagc acactcagtg acatagaaat ctccatcatc
     4021 tgcacccagg ccagcagtca agcacaaggt gcctgtacac tggttctcaa acagacctca
     4081 acctcatctc cttctgtggt cttattatag tcatcaggaa gtggaagtcg gccatctcca
     4141 aaaggaacga acctgtctct cactctgtct gttgttttca atgaagacag tgacagagtt
     4201 ttcatgggca tccttcacaa agcacagttc cacactgcca ccttcagcag cacagagatt
     4261 tccccatgtc ttttgattca gatcaaacca ggatttctgt cttcgataca gaaaataatt
     4321 ttagaagtgg ccatatgaca cagagctgga gattttatta gcgataacgt aacacagatt
     4381 tgaagcatga aaggaaggag aaggagaagc actcgggtag aaacgacagc acacacgggt
     4441 atgtgcttct gaggagaaag ggaccgccgg cgattattac tcttacaacg cgtgcggccg
     4501 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4561 gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4621 ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5101 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5221 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5281 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5341 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5401 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5461 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5521 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5581 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5641 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5701 ttgttgc
//
