LOCUS       sequence-469226-        4813 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 4813)
  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 4813)
  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: AiP1772 - pAAV-AiE2403m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1475..1484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1481..1483,1484..1486,1487..2197)
                     /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(1526..1547)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1787..1806)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2134..2155
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2290..2310)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2497..2608
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2726..2866
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2726..2855
                     /label=AAV2 ITR
     rep_origin      2941..3396
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3028..3047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3238..3259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3413..3432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3532..3554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3592..3610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3678..3782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3783..3851,3852..4643)
                     /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(4001..4020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgttgtaag agtaataatc ggcatgacaa aggggtagca acccacaggt
      841 tgagaactgc tgatgtagag aaatcaaaaa cacggggcaa cttcatccac agacacttca
      901 cagagaattc ttaggaaaca ccaaaggcat ttgtatgaga caacaatcct tcaagtctaa
      961 actcatcatt tcctgaaccc aggctattgt gaatgaggaa aagaagctga taaagggtgg
     1021 ccagttgctt cagccagcaa cgtgcttgtc accatgcctg acaacctgag tttccttccc
     1081 agaaccaacg tagtggaagg aacagactaa ctcccacagg ttttccttac acctccacac
     1141 atgtgttgcg acacatggca ctgttgtgca tctgtgctta cccacatacc taggaacaca
     1201 gtaagtgaat ccaatttaag ggtttttttt tttcaggatc agcaaatcac gggtcaaatc
     1261 gtagctctgc caccgaccgt caccctctga gctagctcct tcaaccctct ctctgccttt
     1321 aacttctact gtctataaag ataatggcaa tatatatccc ataaggatac tggagggatc
     1381 ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     1441 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     1501 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1561 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     1621 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     1681 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1741 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1801 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1861 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1921 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     1981 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     2041 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     2101 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2161 cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc
     2221 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2281 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2341 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     2401 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2461 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     2521 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2581 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2641 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     2701 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     2761 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2821 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     2881 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     2941 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3001 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3061 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3121 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3181 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3241 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3301 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3361 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3421 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3481 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3541 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3601 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3661 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3721 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3781 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3841 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3901 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3961 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4021 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4081 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4141 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4201 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4261 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4321 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4381 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4441 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4501 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4561 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4621 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4681 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4741 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4801 tcaaaggatc ttc
//
