LOCUS       sequence-432591-        5052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2375m 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 5052)
  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 5052)
  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: AiP1920 - pAAV-AiE2375m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   330..459
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(521..1109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(601..620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1280..2071,2072..2140))
                     /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     1903..1922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2141..2245)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2313..2331
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2369..2391)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2491..2510
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2527..2982)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2664..2685)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2876..2895
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3057..3197
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3068..3197
                     /label=AAV2 ITR
     polyA_signal    3315..3426
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3613..3633
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3726..4436,4437..4439,4440..4442))
                     /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(3768..3789)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4117..4136
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4376..4397
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4439..4448
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ttgacgtcta tgcgttttag ccatggagac gacagagcaa acgtcatccc tccctcctcc
       61 tcctcctcct cgcctccccg ccctccctct gcgccagggg cggaggagcg tcgccatcac
      121 gtgttgtcag ctgggctgga ggtctggtgg gcggggcctg ttgtcatgac tgccgaggct
      181 gctgttgttg tttggagctg taattagaaa gtgctgaggc ggcagtggag ccagaacaga
      241 ttctggaaca gtgtctaccg cccccgggcc agcgagggtt ccccagccct gggaaatcca
      301 ggacaccccc taggctgtcc tgcggccgca ggaaccccta gtgatggagt tggccactcc
      361 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
      421 cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc
      481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
      961 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1021 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1201 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1261 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1381 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1441 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1561 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1801 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2161 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2221 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     2281 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     2341 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     2401 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     2461 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     2521 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2581 gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc
     2641 ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2701 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2761 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2821 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2881 agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2941 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
     3001 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg
     3061 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
     3121 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     3181 catcactagg ggttcctgcg gccgctcggt ccgcacgtga gatctcatgc ctgctattgt
     3241 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
     3301 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
     3361 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
     3421 gcacagtcga agatctctcg agccacggaa ttgtcagtgc ccaacagccg agcccctgtc
     3481 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caagaactaa ccaggattta
     3541 tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa
     3601 gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca
     3661 caaattttgt aatccagagg ttgattatga tatcgaattc gcggccgcgg cgcgccgtcg
     3721 acttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     3781 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg gtagctcagg
     3841 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     3901 tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg
     3961 ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc
     4021 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     4081 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     4141 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     4201 tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc cgtagcccag ggtggtcacg
     4261 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg
     4321 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     4381 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     4441 atggtggcga ccggtagcgc tagcttcgaa agatctggat cccagaagca aatgtaagca
     4501 atagatggct ctgccctgac ttttatgccc agcccgagct ccgcaggaat tgggtatgaa
     4561 cggctccgac tgcttttgag agcatgaaaa gcattaactg ggaagccaac ccatttctag
     4621 gacttggctg agaagataat ggcattgctg agcggagatg cgggctggga gccgacgtcc
     4681 tgacactaaa ataccgagct gtaacagaca ctgagaacga ggagcaagac tcccacagga
     4741 gacttggtgt gactatggag cgatgtcatc tgtggggcta taatcagtag gagcagagct
     4801 gcacgttgag cacgttttct cctggatctg gttcttggcg agggatccag accgcagcct
     4861 ccaagatctg ttacagaaat aaacagctgg gagacaacaa ccaaaacagc aacacgggca
     4921 gagcgggcgg gagccgaagc tccgcccctg ctagcctccg accaatcaga agcctgttgc
     4981 cggggtcctg tgttgggcag cctgacgttc ccatagagac caaagaacat tacacactaa
     5041 acactggggg cg
//
