LOCUS       sequence-470883-        4976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0674m_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 4976)
  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 4976)
  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: AiP15081 -
            pAAV-AiE0674m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4976
                     /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      988..997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(994..996,997..999,1000..1710)
                     /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(1039..1060)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1300..1319)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1647..1668
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1803..1823)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2010..2121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2239..2379
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2239..2368
                     /label=AAV2 ITR
     rep_origin      2454..2909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2541..2560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2751..2772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2926..2945)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3045..3067
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3105..3123)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3191..3295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3296..3364,3365..4156)
                     /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(3514..3533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4327..4915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4816..4835
                     /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 gcgtgctgtt cattttaagg aaaggcttat gatatttcag
      181 ctctaacagt ttatggttca gttttaaaat aacagaaatt catagttaag atgttaatgc
      241 attctttgca gctttttaga tatgtataag tataatttta gtttaagaaa atgataaata
      301 tagatttgat ttaaggattc tcctcaattt gcctcacagc ctccctttta gatggcaggt
      361 ttctaaagac tggtggtatt atctctgata tggtgctgtt cattttaagg aaaggcttat
      421 gatatttcag ctctaacagt ttatggttca gttttaaaat aacagaaatt catagttaag
      481 atgttaatgc attctttgca gctttttaga tatgtataag tataatttta gtttaagaaa
      541 atgataaata tagatttgat ttaaggattc tcctcaattt gcctcacagc ctccctttta
      601 gatggcaggt ttctaaagac tggtggtatt atctctgata tggtgctgtt cattttaagg
      661 aaaggcttat gatatttcag ctctaacagt ttatggttca gttttaaaat aacagaaatt
      721 catagttaag atgttaatgc attctttgca gctttttaga tatgtataag tataatttta
      781 gtttaagaaa atgataaata tagatttgat ttaaggattc tcctcaattt gcctcacagc
      841 ctccctttta gatggcaggt ttctaaagac tggtggtatt atctctgata tggtgagctc
      901 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
      961 gatctttcga agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
     1021 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1081 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     1141 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg
     1201 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1261 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1321 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1381 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1441 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1501 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     1561 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     1621 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1681 atcactctcg gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt
     1741 cgatatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1801 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1861 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg
     1921 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     1981 gacaattccg tggctcgaga gatcttcgac tgtgccttct agttgccagc catctgttgt
     2041 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2101 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2161 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg agatctcacg
     2221 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2281 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2341 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     2401 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     2461 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2521 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2581 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2641 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     2701 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2761 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     2821 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2881 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     2941 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3001 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3061 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3121 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3181 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3241 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3301 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3361 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3421 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3481 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3541 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3601 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3661 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3721 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3781 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3841 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3901 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3961 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4021 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4081 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4141 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4201 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4261 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4321 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4381 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4441 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4501 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4561 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4621 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4681 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4741 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4801 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4861 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4921 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
