LOCUS       sequence-466258-        4980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2139m 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 4980)
  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 4980)
  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: AiP1428 - pAAV-AiE2139m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..35)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(695..1486,1487..1555))
                     /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     1318..1337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1556..1660)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1728..1746
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1784..1806)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1906..1925
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1942..2397)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2079..2100)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2291..2310
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2472..2612
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2483..2612
                     /label=AAV2 ITR
     polyA_signal    2730..2841
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3028..3048
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3141..3851,3852..3854,3855..3857))
                     /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(3183..3204)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3532..3551
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3791..3812
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3854..3863
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4725..4854
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4916..4980,1..524))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
       61 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      121 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      181 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      241 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      301 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      361 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      421 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      481 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      541 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      601 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      661 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      721 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      781 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      841 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      901 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      961 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1021 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1081 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1141 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1201 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1261 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1321 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1381 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1441 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1501 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1561 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1621 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1681 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1741 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1801 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1861 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     1921 ttgtactgag agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa
     1981 atttttgtta aatcagctca ttttttaacc aatagaccga aatcggcaaa atcccttata
     2041 aatcaaaaga atagcccgag atagagttga gtgttgttcc agtttggaac aagagtccac
     2101 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2161 cactacgtga accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2221 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2281 cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg
     2341 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact
     2401 atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2461 catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
     2521 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     2581 gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtgagatct
     2641 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     2701 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     2761 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     2821 gctggcaact agaaggcaca gtcgaagatc tctcgagcca cggaattgtc agtgcccaac
     2881 agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaaga
     2941 actaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga
     3001 tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata
     3061 ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatgatatcg aattcgcggc
     3121 cgcggcgcgc cgtcgactta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     3181 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg
     3241 gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     3301 ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg
     3361 aagttggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt gtggctgttg
     3421 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     3481 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     3541 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     3601 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg cacgccgtag
     3661 cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatcagc
     3721 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     3781 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     3841 tcgcccttgc tcaccatggt ggcgaccggt agcgctagct tcgaaagatc tggatcccag
     3901 aagcaaatgt aagcaataga tggctctgcc ctgactttta tgcccagccc gagctccgcc
     3961 ggcgttcggt gaatgaacca ggaagtgaga gaaagctgtg ttctgctacc taaaacctca
     4021 ccgtctgtat ttcacacagg caaaatgcat gtgaaaccaa ttcccaggca ctcaccacgt
     4081 gtgcatcaca gcggtgcgcc ccccagagag ctcccatccc tccatccagg aattacagtt
     4141 ccttttcatt ggctccgccc ccaaagagct cttatccctg cgtccatgag ttacagttcg
     4201 ttttcattgg ctcatgtccc tgctcaagcc tgaaagtcag tggtgtgagc agtgccaggc
     4261 taaaagtgga ctggaaagaa tttgctagag tatacaagtc taataatttt ccggatgtga
     4321 ggcgctgtgc tcgctttgcc gccctgcctg tccgtcttcg cgggctgctc ctgaagccct
     4381 ctgggagaca ttagtccttt gtcttgactg gggcagggct cagtgaccaa gcccaggtgg
     4441 gctagttgcg tatttaatcc aagggtggtg ttatcattga gaccttagtt gtccccccaa
     4501 gacctccctg ctaaagacat tgtccccagg atgtactgga aggtcctgga acctccagga
     4561 ggtgggaccc gctgtgagga acttagatca ctgggggatg ctcttcaaga atctcagagg
     4621 gtcaacaagg aacacgctaa acacattcca tgtggtattc acagactttg tgacgtcttt
     4681 cctggggcgc cggcgattat tactcttaca acgcgtgcgg ccgcaggaac ccctagtgat
     4741 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4801 cgcccgacgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggacatgt
     4861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
//
