LOCUS       sequence-466340-        4526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2609m 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 4526)
  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 4526)
  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: AiP2037 - pAAV-AiE2609m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      285..294
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(291..293,294..296,297..1007)
                     /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(336..357)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(597..616)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     944..965
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1100..1120)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1307..1418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1536..1676
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1536..1665
                     /label=AAV2 ITR
     rep_origin      1751..2206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1838..1857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2048..2069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2223..2242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2342..2364
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2402..2420)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2488..2592
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2593..2661,2662..3453)
                     /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(2811..2830)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3624..4212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4113..4132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4274..4403
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ccccagatat gctccccaag tagaatattc aggtccctac agacccgcac gcagagagct
       61 gacaattccc gcgacaggca acctaagcgg tcccgcttca tctccctagc gggactcgac
      121 ggtcggggat agagagaggg ctccgagggt gctcactctt gactgttccc ttctccccgc
      181 agacgccggc ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca
      241 tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca
      301 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      361 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      421 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      481 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      541 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      601 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      661 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      721 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
      781 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
      841 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
      901 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      961 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg
     1021 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     1081 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1141 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     1201 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1261 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     1321 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1381 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1441 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     1501 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     1561 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     1621 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     1681 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     1741 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1801 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     1861 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1921 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     1981 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2041 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     2101 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     2161 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     2221 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     2281 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2341 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     2401 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2461 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2521 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2581 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2641 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2701 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2761 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2821 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2881 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2941 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3001 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3061 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3121 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3181 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3241 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3301 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3361 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3421 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3481 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3541 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3601 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3661 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3721 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3781 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3841 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3901 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3961 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4021 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4081 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4141 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4201 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4261 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
     4321 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
     4381 caactccatc actaggggtt cctgcggccg cacgcgtatg ggtctatgga agccactggg
     4441 accttgcggt cccgcacacc tgcaccggga gcgggaaaaa cctcgctcgc ctgcctccct
     4501 cccagagcgc gcagttatcc cgcggg
//
