LOCUS       sequence-466194-        4784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2368m 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 4784)
  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 4784)
  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: AiP1693 - pAAV-AiE2368m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      286..295
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(292..294,295..297,298..1008)
                     /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(337..358)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(598..617)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     945..966
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1101..1121)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1308..1419
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1537..1677
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1537..1666
                     /label=AAV2 ITR
     rep_origin      1752..2207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1839..1858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2049..2070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2224..2243)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2343..2365
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2403..2421)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2489..2593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2594..2662,2663..3454)
                     /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     complement(2812..2831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3625..4213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4114..4133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4275..4404
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tctgttgggc tgtgatgagg tagctacgag agacggtgcg gggaggaaga acactgaatg
       61 gccagaggcc cagagggctt tctgttgtga tccagcccca gatgctgatg atctggcctc
      121 ttagagtggg ggatggaggg atcatacccg ctcgggtaga gggaagaagt gggatcaccc
      181 tggaagactt cggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
      241 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc
      301 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      361 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      421 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      481 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
      541 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      601 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      661 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      721 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
      781 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
      841 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
      901 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
      961 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc
     1021 gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga
     1081 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1141 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1201 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1261 gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag
     1321 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1381 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1441 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1501 caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     1561 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     1621 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     1681 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     1741 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1801 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     1861 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1921 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     1981 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2041 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     2101 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     2161 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     2221 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2281 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2341 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2401 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2461 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2521 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2581 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2641 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2701 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2761 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2821 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2881 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2941 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3001 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3061 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3121 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3181 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3241 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3301 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3361 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3421 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3481 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3541 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3601 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3661 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3721 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3781 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3841 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3901 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3961 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4021 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4081 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4141 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4201 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4261 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     4321 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     4381 ccaactccat cactaggggt tcctgcggcc gcacgcgttg taagagtaat aatcggaaaa
     4441 atacaaagaa atgagcacaa acacagagga ggtatagagg ctggtgatgg ggaagggaga
     4501 gcaggttgtc taggggagtc caccacacac cttgttgatg agagctgagg ctccctagcc
     4561 atcggatctg ggaacatgtc ccgattcagt ggagcttggc aggccagagc tgtttggtgg
     4621 attgtagtct ggctgttgtc ttccacatta aacataattg gctgcaaata tctataccag
     4681 tgcaacacac attgctttgc aataatttgg ttaaaacaca tttccatcaa agtgcagtca
     4741 acaagaacaa accccacaat gaaccatcat caacctgctg catc
//
