LOCUS       sequence-466237-        5163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2100m 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 5163)
  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 5163)
  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: AiP1286 - pAAV-AiE2100m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     361..380
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   522..651
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1696..1705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1702..1704,1705..1707,1708..2418)
                     /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(1747..1768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2008..2027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2355..2376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2511..2531)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2718..2829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2947..3087
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2947..3076
                     /label=AAV2 ITR
     rep_origin      3162..3617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3249..3268)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3459..3480
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3634..3653)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3753..3775
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3813..3831)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3899..4003
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4004..4072,4073..4864)
                     /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(4222..4241)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5035..5163,1..460)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
       61 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      121 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      181 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      241 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      301 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      361 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      421 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      481 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
      541 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
      601 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
      661 cgcgtcgtca aacacagctc aagctgacct tgatttgctt gtaatcctcc tattttggct
      721 ttctggatgc taggacaaag gcatgagcca ccagccctag taagattagg ctttgttttg
      781 tttgtatttc cttgacttag caattagaaa gcctaaaaga aaccacttgt gatcaaattc
      841 aatgtgcttc caaagttcgc tgaggatcag atgtcgtggg gatggcagac ttcaaacctg
      901 tttctgaact attttgctgt gtctttatgt cccaggtcaa aggatttcaa ctgatgctcc
      961 ataaagcaag aagggaaaat ggatttctta atgagtctgt atacagtgca gtatctatga
     1021 gctatggtgt gtgtgactgc caaggcagtc agatgctatt caagcaagga atgctgctct
     1081 gagatcagct agtacgtaag aaagacatcc tggctcctgg aagatgacaa gagtttgcct
     1141 ccgtagtatt tgaaagaaag tggtcatggc catcatgaat caatcacttc aggaaaaggt
     1201 tgagcatgtg ctgtgacgac tgcagatctt tttatagctg tggattcttt tatagctgct
     1261 gagatacgac gataggagtg gagcggtagt tatcgttgat ttgtttggta atgtttcagg
     1321 caggtctacg aattcctttg gattgtgggt tgtctcccaa tcctacactt tcctagctct
     1381 ctaagaattc ccagagggta tggagccagt agagactgac cacaaaagtt gacacttaat
     1441 gttggctgac ctgcaaagaa ggtttcatta tgcattcttt tctgcacatg tgaaatcaga
     1501 gaccatggga tccatttcaa taacaaagcc catctttgta aagtagaagc tagttgaaaa
     1561 ttaagtatca ttttctattg tggtgattgg gaagaaatat tggagctcgg gctgggcata
     1621 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
     1681 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1741 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1801 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     1861 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     1921 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1981 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2041 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2101 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2161 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     2221 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2281 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     2341 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2401 atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat
     2461 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2521 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2581 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     2641 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2701 gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2761 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2821 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2881 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     2941 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3001 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3061 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3121 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3181 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     3241 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3301 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3361 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3421 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3481 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3541 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3601 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3661 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3721 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3781 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3841 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3901 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3961 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4021 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4081 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4141 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4201 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4261 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4321 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4381 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4441 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4501 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4561 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4621 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4681 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4741 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4801 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4861 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4921 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4981 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5041 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5101 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5161 agc
//
