LOCUS       sequence-452926-        5136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0354m_3xC3 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 5136)
  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 5136)
  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: AiP12566 -
            pAAV-3xSP10ins-AiE0354m_3xC3-minRho...
FEATURES             Location/Qualifiers
     source          1..5136
                     /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      1148..1157
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1154..1156,1157..1159,1160..1870)
                     /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(1199..1220)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1460..1479)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1807..1828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1963..1983)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2170..2281
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2399..2539
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2399..2528
                     /label=AAV2 ITR
     rep_origin      2614..3069
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2701..2720)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2911..2932
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3086..3105)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3205..3227
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3265..3283)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3351..3455
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3456..3524,3525..4316)
                     /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(3674..3693)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4487..5075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4976..4995
                     /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 cgcgccggta ccgaagctac ccctaacaca
      181 ctattctaca cacagaaaat gctcttcact aggaagctac ccctaacaca ctattctaca
      241 cacagaaaat gctcttcact aggaagctac ccctaacaca ctattctaca cacagaaaat
      301 gctcttcact agacgcgtga cctttaacaa tgaatcttta aatttaaccg ggtcccccag
      361 agatcaccgc acacaaggtt tcattgtgtt tcgatctgag gtgagattca aatgtgggcc
      421 gtgggattcc tgttttgggt agagcggtta acacagaaca gtaggatagg tcagcaatta
      481 tcagccaaga acaaaggaga accctgatgc tcagtctacc ttattatggg gtctcacttt
      541 ggacctttaa caatgaatct ttaaatttaa ccgggtcccc cagagatcac cgcacacaag
      601 gtttcattgt gtttcgatct gaggtgagat tcaaatgtgg gccgtgggat tcctgttttg
      661 ggtagagcgg ttaacacaga acagtaggat aggtcagcaa ttatcagcca agaacaaagg
      721 agaaccctga tgctcagtct accttattat ggggtctcac tttggacctt taacaatgaa
      781 tctttaaatt taaccgggtc ccccagagat caccgcacac aaggtttcat tgtgtttcga
      841 tctgaggtga gattcaaatg tgggccgtgg gattcctgtt ttgggtagag cggttaacac
      901 agaacagtag gataggtcag caattatcag ccaagaacaa aggagaaccc tgatgctcag
      961 tctaccttat tatggggtct cactttggag ctcgattcag ccgggagctt agggagggga
     1021 ggtcacttca taagggcttg gggggggagt tggagccacg agtcgtccag ccggagcccc
     1081 gtgtggctgt gctccggcct cagaagcatc cccggatcca gatctttcga agctagcgct
     1141 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1201 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1261 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
     1321 gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc
     1381 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1441 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1501 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1561 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga
     1621 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
     1681 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1741 gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg
     1801 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1861 gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct
     1921 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1981 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2041 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     2101 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga
     2161 gatcttcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2221 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2281 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2341 gggaggattg ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag
     2401 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     2461 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     2521 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2581 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     2641 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2701 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2761 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2821 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2881 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2941 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3001 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     3061 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3121 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3181 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3241 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3301 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3361 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3421 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3481 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3541 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3601 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3661 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3721 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3781 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3841 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3901 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3961 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4021 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4081 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4141 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4201 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4261 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4321 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4381 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4441 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4501 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4561 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4621 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4681 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4741 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4801 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4861 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4921 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4981 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5041 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5101 ggttcctggc cttttgctgg ccttttgctc acatgt
//
