LOCUS       sequence-438957-        4542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2340m 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 4542)
  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 4542)
  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: AiP1658 - pAAV-AiE2340m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     162..180
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(218..240)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     340..359
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(376..831)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(513..534)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     725..744
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   906..1046
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   917..1046
                     /label=AAV2 ITR
     polyA_signal    1164..1275
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1462..1482
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1575..2285,2286..2288,2289..2291))
                     /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(1617..1638)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1966..1985
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2225..2246
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2288..2297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2721..2850
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2912..3500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2992..3011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3671..4462,4463..4531))
                     /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     4294..4313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4532..4542,1..94))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
       61 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
      121 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
      181 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      241 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      301 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      361 tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
      421 gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct tataaatcaa
      481 aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt ccactattaa
      541 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
      601 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
      661 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
      721 aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc
      781 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt
      841 gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      901 gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
      961 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     1021 ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgag atctcatgcc
     1081 tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
     1141 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
     1201 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
     1261 aactagaagg cacagtcgaa gatctctcga gccacggaat tgtcagtgcc caacagccga
     1321 gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aagaactaac
     1381 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     1441 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     1501 aatctttcac aaattttgta atccagaggt tgattatgat atcgaattcg cggccgcggc
     1561 gcgccgtcga cttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg
     1621 aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg
     1681 tagctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc
     1741 tggtagtggt cggcgagctg cacgccgccg tcctcgatgt tgtggcggat cttgaagttg
     1801 gccttgatgc cgttcttctg cttgtcggcg gtgatataga cgttgtggct gttgtagttg
     1861 tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg
     1921 atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg
     1981 tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag
     2041 aagtcgtgct gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc gtagcccagg
     2101 gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat cagcttcagg
     2161 gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg
     2221 tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc
     2281 ttgctcacca tggtggcgac cggtagcgct agcttcgaaa gatctggatc ccagaagcaa
     2341 atgtaagcaa tagatggctc tgccctgact tttatgccca gcccgagctc cggatgagga
     2401 gagacaagag gaagtaagaa aggatgggaa gaaactgctt tggttggtgg gaaaaggtca
     2461 ttatcttaat ttgtgatgat gatttatggc tgtacacatg tgtcagagct cattaaatca
     2521 tacacatcac aatgtgctgt tgattgtatg tcaactatac ttcaataaat ctgaaaaaaa
     2581 tgaattcact gggatgagtg ccatgagaaa ggagctctga aatagtttgt ggaagcagtg
     2641 agatggaact tatccagact tatgctgaga catagggaga tatcacacgc gattattact
     2701 cttacaacgc gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2761 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag
     2821 tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca
     2881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4501 taagggcgac acggaaatgt tgaatactca tactcttcct tt
//
