LOCUS       sequence-449772-        5554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0476m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical Chandelier cells (note
            HPF ChC cells).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  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 5554)
  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: AiP12782 -
            pAAV-AiE0476m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      1566..1575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1572..1574,1575..1577,1578..2288)
                     /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-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1878..1897)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2225..2246
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2381..2401)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2588..2699
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2817..2957
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2817..2946
                     /label=AAV2 ITR
     rep_origin      3032..3487
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3119..3138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3329..3350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3504..3523)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3623..3645
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3683..3701)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3769..3873
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3874..3942,3943..4734)
                     /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(4092..4111)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4905..5493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5394..5413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtagaaa caccacaagc aaagccaaaa
      181 taggtggcct agaacttcca gcttgaaata tgggagagaa tgagggaggc actgtagagc
      241 agctgccggg tgccgcatga gaacaattct ccctgctcat aattaatcct acctatttct
      301 gatgacagct ggctcttcac tttgaacaag ctagttaaca actttcttct cacattgagc
      361 aaataattca tatttaatta cttaaccacc agttacaaaa tgagaatcat caaggaatca
      421 caattaattt gctattgaca aactcatact tttagcaggc tgatttctac tttatactta
      481 gattggtaat gaaaaatgaa gcttatttta gttgattggt tggacttgtg tatgaatatt
      541 atctattatt tgaaaagcca aacttgaatg caaaaaaata ttgaatatga aaagagaaac
      601 accacaagca aagccaaaat aggtggccta gaacttccag cttgaaatat gggagagaat
      661 gagggaggca ctgtagagca gctgccgggt gccgcatgag aacaattctc cctgctcata
      721 attaatccta cctatttctg atgacagctg gctcttcact ttgaacaagc tagttaacaa
      781 ctttcttctc acattgagca aataattcat atttaattac ttaaccacca gttacaaaat
      841 gagaatcatc aaggaatcac aattaatttg ctattgacaa actcatactt ttagcaggct
      901 gatttctact ttatacttag attggtaatg aaaaatgaag cttattttag ttgattggtt
      961 ggacttgtgt atgaatatta tctattattt gaaaagccaa acttgaatgc aaaaaaatat
     1021 tgaatatgaa aagagaaaca ccacaagcaa agccaaaata ggtggcctag aacttccagc
     1081 ttgaaatatg ggagagaatg agggaggcac tgtagagcag ctgccgggtg ccgcatgaga
     1141 acaattctcc ctgctcataa ttaatcctac ctatttctga tgacagctgg ctcttcactt
     1201 tgaacaagct agttaacaac tttcttctca cattgagcaa ataattcata tttaattact
     1261 taaccaccag ttacaaaatg agaatcatca aggaatcaca attaatttgc tattgacaaa
     1321 ctcatacttt tagcaggctg atttctactt tatacttaga ttggtaatga aaaatgaagc
     1381 ttattttagt tgattggttg gacttgtgta tgaatattat ctattatttg aaaagccaaa
     1441 cttgaatgca aaaaaatatt gaatatgaaa aggagctcgg gctgggcata aaagtcaggg
     1501 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
     1561 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1621 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1681 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1741 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1801 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1861 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1921 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1981 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     2041 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     2101 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2161 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     2221 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2281 tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg
     2341 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2401 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2461 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     2521 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagaga
     2581 tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2641 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2701 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2761 gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc ggccgcagga
     2821 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     2881 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     2941 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3001 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3061 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3121 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3181 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3241 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3301 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3361 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3421 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3481 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3541 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3601 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3661 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     3721 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3781 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3841 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3901 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3961 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4021 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4081 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4141 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4201 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4261 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4321 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4381 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4441 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4501 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4561 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4621 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4681 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4741 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4801 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4861 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4921 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4981 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5041 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5101 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5161 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5221 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5281 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5341 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5401 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5461 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5521 ttcctggcct tttgctggcc ttttgctcac atgt
//
