LOCUS       sequence-466284-        4748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2341m 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 4748)
  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 4748)
  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: AiP1659 - pAAV-AiE2341m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(579..1370,1371..1439))
                     /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     1202..1221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1440..1544)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1612..1630
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1668..1690)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1790..1809
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1826..2281)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1963..1984)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2175..2194
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2356..2496
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2367..2496
                     /label=AAV2 ITR
     polyA_signal    2614..2725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2912..2932
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3025..3735,3736..3738,3739..3741))
                     /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(3067..3088)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3416..3435
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3675..3696
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3738..3747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4377..4506
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4568..4748,1..408))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4648..4667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
       61 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      121 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      181 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      241 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      301 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      361 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      421 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      481 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      541 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
      601 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      661 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
      721 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
      781 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
      841 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
      901 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
      961 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1021 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1081 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1141 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     1201 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1261 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1321 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1381 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1441 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1501 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     1561 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     1621 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     1681 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     1741 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     1801 cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg
     1861 ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct
     1921 tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt
     1981 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2041 ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca
     2101 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2161 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta
     2221 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2281 tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2341 accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     2401 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     2461 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgag
     2521 atctcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
     2581 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
     2641 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
     2701 gatggctggc aactagaagg cacagtcgaa gatctctcga gccacggaat tgtcagtgcc
     2761 caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc
     2821 aagaactaac caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc
     2881 atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag
     2941 aataccagtc aatctttcac aaattttgta atccagaggt tgattatgat atcgaattcg
     3001 cggccgcggc gcgccgtcga cttacttgta cagctcgtcc atgccgagag tgatcccggc
     3061 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     3121 cttggactgg tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     3181 ggtgttctgc tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcggat
     3241 cttgaagttg gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtggct
     3301 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     3361 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     3421 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     3481 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc
     3541 gtagcccagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     3601 cagcttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     3661 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     3721 ctcctcgccc ttgctcacca tggtggcgac cggtagcgct agcttcgaaa gatctggatc
     3781 ccagaagcaa atgtaagcaa tagatggctc tgccctgact tttatgccca gcccgagctc
     3841 cgtgcctctt cacagcaaca gtaccataac taagacagca atcagcttca tataagttca
     3901 acatagtttc tattctgatt aatctctgaa caataccaca ttcacccctt tagcaatatt
     3961 gcatttaaaa taatgagttt tcatgaacct ggtgaaatta ttactgacag tgtttcttct
     4021 cactcttaac cttttccaca ctactctttc caagactcaa gtggaaaatt ctcttagttt
     4081 atattcttgt aagatgctgg gcttacataa acttgtgctc cctgaaaaat gtcagagaga
     4141 taatttaatg caagagcctt tctcaggcag aacatcagga taaataattt taaaaacaca
     4201 ttgaaaggat acccgaatcc cgtagttcca gctggataat aaatcacaaa attaagtctt
     4261 atttctgaca ttaacaaatg aaattccatt aggctgggaa gagaactgct atttagcttt
     4321 cagccatata tgtaaataat gggtacgatt attactctta caacgcgtgc ggccgcagga
     4381 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4441 gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4501 tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4561 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4621 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4681 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4741 cgggaagc
//
