LOCUS       sequence-461967-        4974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3050m 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 4974)
  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 4974)
  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: AiP20016 - pAAV-AiE3050m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      986..995
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(992..994,995..997,998..1708)
                     /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(1037..1058)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1298..1317)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1645..1666
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1801..1821)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2008..2119
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2237..2377
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2237..2366
                     /label=AAV2 ITR
     rep_origin      2452..2907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2539..2558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2749..2770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2924..2943)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3043..3065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3103..3121)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3189..3293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3294..3362,3363..4154)
                     /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(3512..3531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4325..4913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4814..4833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatacca cgtacctgcc ataatcaata ctgcttagaa
      181 tataaactgc aggatccttc tcagcttaca ggaaaataaa taaagcaaaa gagacactct
      241 ggatcccagg gggattcttt tcccaaagca atcagacatc tggagtcttg gtctgataaa
      301 aatcaatggc tctttcaggg cagtacatct tttagatgcg actgaggaaa gcagtgcata
      361 atgatgacag aactggagga gggaaaggca agcaaagaaa aacaatcata cgtaaaactc
      421 taaccttttt tgctgtaata gactgtgcca aaacatagtc tgtgggataa ttacatgcat
      481 atatttatag gactctgaat cttgaaatgg tagtgaaaca cagttcttta gagtctggct
      541 cttgtgggta cagtctttac tttctctttt gtatatctgt ctgtgcttgt gtgttatgat
      601 ccaaatgtat aggaactatc acattttatt cagtagtaag tatagtgtgc tatgtgcaca
      661 taattgcagg aaaacactac gaggcagttt gagtcatgag ctcctaagat agccgttcta
      721 atggacactg aatctgaact gagagggggc agggcccagg gccgctcagc aacagcacag
      781 atgacctttt ctgtattgat cactctatgc tcgcagtcac ccactcaaaa aaatattgca
      841 gcaagccaag tacttatata cattcaggca gaagactact catgcgccgg cggagctcgg
      901 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
      961 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1021 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1081 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     1141 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
     1201 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1261 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1321 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1381 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1441 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     1501 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1561 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     1621 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1681 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     1741 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1801 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1861 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     1921 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     1981 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2041 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2101 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2161 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     2221 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     2281 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     2341 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     2401 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2461 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2521 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2581 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2641 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     2701 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2761 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     2821 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2881 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     2941 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3001 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3061 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3121 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3181 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3241 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3301 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3361 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3421 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3481 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3541 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3601 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3661 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3721 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3781 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3841 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3901 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3961 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4021 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4081 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4141 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4201 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4261 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4321 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4381 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4441 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4501 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4561 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4621 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4681 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4741 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4801 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4861 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4921 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
