LOCUS       sequence-466361-        4707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE3094m 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 4707)
  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 4707)
  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: AiP20051 - pAAV-AiE3094m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      248..257
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(254..256,257..259,260..970)
                     /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(299..320)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(560..579)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     907..928
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1063..1083)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1270..1381
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1499..1639
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1499..1628
                     /label=AAV2 ITR
     rep_origin      1714..2169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1801..1820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2011..2032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2186..2205)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2305..2327
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2365..2383)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2451..2555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2556..2624,2625..3416)
                     /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(2774..2793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3587..4175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4076..4095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4237..4366
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tggccatagc taacgcatgc tgcgctcctc gattccccac cctgggcggc cgcgtggtca
       61 atgtggcgca gatcatcgag ctggtgatga cgtgtatctt gtacgtcgtg gtgagcggca
      121 acctcatgta caacagtttc ccggggcgcc ggcggagctc gggctgggca taaaagtcag
      181 ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct
      241 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      301 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      361 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
      421 gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc
      481 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      541 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      601 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      661 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga
      721 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
      781 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      841 gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg
      901 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
      961 gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct
     1021 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1081 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     1141 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     1201 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga
     1261 gatcttcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1321 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1381 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     1441 gggaggattg ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag
     1501 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     1561 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     1621 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     1681 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     1741 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     1801 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1861 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1921 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1981 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2041 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     2101 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     2161 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2221 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2281 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2341 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     2401 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2461 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2521 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2581 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2641 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2701 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2761 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2821 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2881 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2941 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3001 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3061 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3121 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3181 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3241 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3301 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3361 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3421 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3481 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3541 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3601 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3661 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3721 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3781 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3841 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3901 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3961 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4021 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4081 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4141 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4201 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
     4261 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     4321 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
     4381 ctttcacgag gattcacccc cattctctcg tcctgtggct taggcacaag tctgtctcca
     4441 tctagcgccc cctagcgata gttttgggct ataccccagg gtcccttttc aagaggatca
     4501 gctgagctcc tgggtctggt tgcctctttg caccacaggg catgttcgtg ctgggcctac
     4561 cctacgccat cctccacggc ggctacctgg ggttgttcct catcatcttc gccgcagtgg
     4621 tgtgctgcta caccggcaag atcctcatcg cgtgcctgta cgaggagaac gaagacgggg
     4681 aggtggtgcg cgtgcgggac tcgtatg
//
