LOCUS       sequence-460605-        4844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0359h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4844)
  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 4844)
  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: AiP12780 -
            pAAV-AiE0359h_3xC2b-minBG-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..4844
                     /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      856..865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(862..864,865..867,868..1578)
                     /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(907..928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1168..1187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1515..1536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1671..1691)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1878..1989
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2107..2247
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2107..2236
                     /label=AAV2 ITR
     rep_origin      2322..2777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2409..2428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2619..2640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2794..2813)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2913..2935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2973..2991)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3059..3163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3164..3232,3233..4024)
                     /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(3382..3401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4195..4783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4684..4703
                     /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 gcgtcctcat ctgggcatta gaagtcattc tgaagaggct
      181 gtcctatatc tcagtgcaat ttcctactta tccactcccg actataccct gctttcatca
      241 gattgcaact gacctaattt gtaaagactt cagtgaaaag agcctcatta aaagccttca
      301 tctcttttgc agcctctctt tcctccctca aatgatccag gtcagaatgg cctcatctgg
      361 gcattagaag tcattctgaa gaggctgtcc tatatctcag tgcaatttcc tacttatcca
      421 ctcccgacta taccctgctt tcatcagatt gcaactgacc taatttgtaa agacttcagt
      481 gaaaagagcc tcattaaaag ccttcatctc ttttgcagcc tctctttcct ccctcaaatg
      541 atccaggtca gaatggcctc atctgggcat tagaagtcat tctgaagagg ctgtcctata
      601 tctcagtgca atttcctact tatccactcc cgactatacc ctgctttcat cagattgcaa
      661 ctgacctaat ttgtaaagac ttcagtgaaa agagcctcat taaaagcctt catctctttt
      721 gcagcctctc tttcctccct caaatgatcc aggtcagaat gggagctcgg gctgggcata
      781 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
      841 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      901 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      961 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     1021 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     1081 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1141 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1201 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1261 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1321 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1381 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1441 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     1501 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1561 atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat
     1621 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1681 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     1741 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     1801 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     1861 gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1921 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1981 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2041 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     2101 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2161 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2221 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     2281 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     2341 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     2401 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2461 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2521 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2581 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2641 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     2701 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2761 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2821 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     2881 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     2941 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3001 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3061 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3241 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3301 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3361 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3421 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3481 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3541 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3601 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3661 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3721 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3781 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3841 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3901 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3961 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4021 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4081 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4141 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4201 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4261 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4321 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4381 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4441 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4501 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4561 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4621 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4741 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4801 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
