LOCUS       sequence-473598-        4676 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 4676)
  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 4676)
  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: AiP15253 -
            pAAV-AiE0025m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4676
                     /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      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(694..696,697..699,700..1410)
                     /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(739..760)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1000..1019)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1347..1368
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1503..1523)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1710..1821
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1939..2079
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1939..2068
                     /label=AAV2 ITR
     rep_origin      2154..2609
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2241..2260)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2451..2472
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2626..2645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2745..2767
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2805..2823)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2891..2995
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2996..3064,3065..3856)
                     /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(3214..3233)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4027..4615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4516..4535
                     /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 gcgtcactag acctatcgaa tatgcagatg caaaggaaag
      181 ttggtctatt cagcctatcc cgcaacaagc caagaaaacc ctggcaccaa tcaggaacaa
      241 caaaaatgga agaaagtatg tgcttaataa tcggtatcat tcctgaagta aatacactag
      301 acctatcgaa tatgcagatg caaaggaaag ttggtctatt cagcctatcc cgcaacaagc
      361 caagaaaacc ctggcaccaa tcaggaacaa caaaaatgga agaaagtatg tgcttaataa
      421 tcggtatcat tcctgaagta aatacactag acctatcgaa tatgcagatg caaaggaaag
      481 ttggtctatt cagcctatcc cgcaacaagc caagaaaacc ctggcaccaa tcaggaacaa
      541 caaaaatgga agaaagtatg tgcttaataa tcggtatcat tcctgaagta aatagagctc
      601 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
      661 gatctttcga agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
      721 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      781 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
      841 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg
      901 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      961 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1021 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1081 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1141 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1201 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     1261 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     1321 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1381 atcactctcg gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt
     1441 cgatatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1501 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1561 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg
     1621 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     1681 gacaattccg tggctcgaga gatcttcgac tgtgccttct agttgccagc catctgttgt
     1741 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1801 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1861 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg agatctcacg
     1921 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     1981 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2041 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     2101 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     2161 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2221 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2281 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2341 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     2401 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2461 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     2521 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2581 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     2641 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2701 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2761 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     2821 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2881 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2941 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3001 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3061 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3121 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3181 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3241 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3301 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3361 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3421 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3481 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3541 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3601 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3661 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3721 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3781 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3841 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3901 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3961 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4021 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4081 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4141 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4201 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4261 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4321 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4381 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4441 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4501 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4561 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4621 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
