LOCUS       sequence-466318-        4832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2403m 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 4832)
  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 4832)
  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: AiP1875 - pAAV-AiE2403m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      438..447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(444..446,447..449,450..1160)
                     /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(489..510)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(750..769)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1097..1118
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1253..1273)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1460..1571
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1689..1829
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1689..1818
                     /label=AAV2 ITR
     rep_origin      1904..2359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1991..2010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2201..2222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2376..2395)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2495..2517
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2555..2573)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2641..2745
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2746..2814,2815..3606)
                     /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(2964..2983)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3777..4365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4266..4285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4427..4556
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tgacaacctg agtttccttc ccagaaccaa cgtagtggaa ggaacagact aactcccaca
       61 ggttttcctt acacctccac acatgtgttg cgacacatgg cactgttgtg catctgtgct
      121 tacccacata cctaggaaca cagtaagtga atccaattta agggtttttt tttttcagga
      181 tcagcaaatc acgggtcaaa tcgtagctct gccaccgacc gtcaccctct gagctagctc
      241 cttcaaccct ctctctgcct ttaacttcta ctgtctataa agataatggc aatatatatc
      301 ccataaggat actggaggga ttaaataaga atgtgtctat atcggagctc gggctgggca
      361 taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga
      421 agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      481 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      541 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
      601 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg
      661 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      721 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      781 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      841 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
      901 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
      961 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1021 ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca
     1081 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1141 gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata
     1201 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     1261 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     1321 tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca
     1381 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     1441 tggctcgaga gatcttcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1501 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     1561 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     1621 gacagcaagg gggaggattg ggaagacaat agcaggcatg agatctcacg tgcggaccga
     1681 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     1741 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     1801 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     1861 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     1921 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1981 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2041 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2101 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2161 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2221 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     2281 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2341 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2401 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2521 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     2581 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2641 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2701 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2761 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2821 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2881 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2941 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3001 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3061 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3121 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3181 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3241 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3301 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3361 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3421 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3481 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3541 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3601 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3661 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3721 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3781 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3841 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3901 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3961 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4021 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4081 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4141 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4201 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4261 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4321 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4381 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
     4441 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc
     4501 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
     4561 ccgcaaaata tttttggaga tggatgcatg acaaaggggt agcaacccac aggttgagaa
     4621 ctgctgatgt agagaaatca aaaacacggg gcaacttcat ccacagacac ttcacagaga
     4681 attcttagga aacaccaaag gcatttgtat gagacaacaa tccttcaagt ctaaactcat
     4741 catttcctga acccaggcta ttgtgaatga ggaaaagaag ctgataaagg gtggccagtt
     4801 gcttcagcca gcaacgtgct tgtcaccatg cc
//
