LOCUS       sequence-470795-        4679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0025m_3xC2 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 4679)
  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 4679)
  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: AiP15306 -
            pAAV-AiE0025m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4679
                     /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      691..700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(697..699,700..702,703..1413)
                     /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(742..763)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1003..1022)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1350..1371
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1506..1526)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1713..1824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1942..2082
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1942..2071
                     /label=AAV2 ITR
     rep_origin      2157..2612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2244..2263)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2454..2475
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2629..2648)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2748..2770
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2808..2826)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2894..2998
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2999..3067,3068..3859)
                     /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(3217..3236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4030..4618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4519..4538
                     /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 gcgttattta cttcagagaa tgataccgat tattaagcac
      181 atactttctt ccatttttgt tgttcctgat tggtgccagg gttttcttgg cttgttgcgg
      241 gataggctga atagaccaac tttcctttgc atctgcatat tcgataggtc tagtgtattt
      301 acttcagaga atgataccga ttattaagca catactttct tccatttttg ttgttcctga
      361 ttggtgccag ggttttcttg gcttgttgcg ggataggctg aatagaccaa ctttcctttg
      421 catctgcata ttcgataggt ctagtgtatt tacttcagag aatgataccg attattaagc
      481 acatactttc ttccattttt gttgttcctg attggtgcca gggttttctt ggcttgttgc
      541 gggataggct gaatagacca actttccttt gcatctgcat attcgatagg tctagtggag
      601 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
      661 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
      721 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      781 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
      841 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
      901 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      961 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1021 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1081 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1141 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     1201 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     1261 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     1321 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1381 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     1441 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     1501 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     1561 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     1621 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1681 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     1741 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1801 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1861 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     1921 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     1981 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2041 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2101 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     2161 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2221 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2281 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2341 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     2401 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2461 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     2521 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2581 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     2641 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     2701 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2761 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     2821 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     2881 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2941 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3001 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3061 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3121 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3181 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3241 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3301 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3361 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3421 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3481 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3541 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3601 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3661 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3721 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3781 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3841 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3901 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3961 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4021 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4081 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4141 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4201 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4261 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4321 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4381 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4441 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4501 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4561 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4621 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
