LOCUS       sequence-461947-        5040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2069m 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 5040)
  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 5040)
  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: AiP1284 - pAAV-AiE2069m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1052..1061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1058..1060,1061..1063,1064..1774)
                     /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(1103..1124)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1364..1383)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1711..1732
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1867..1887)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2074..2185
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2303..2443
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2303..2432
                     /label=AAV2 ITR
     rep_origin      2518..2973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2605..2624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2815..2836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2990..3009)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3109..3131
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3169..3187)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3255..3359
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3360..3428,3429..4220)
                     /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(3578..3597)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4391..4979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4880..4899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcagcag ggaatagtct caaaatggat tcagggtgaa
      181 ggatgattaa cagtatcagt cagtcaagcc gagggccaga tgatcaaaga ggaatagaat
      241 ttcagtacaa agcaaggcaa agatgctgta tgtgctgata gaagcacaga gggcctggct
      301 ggacaaagtg cagaaagaca atgagctgga caaacgagag actagaatta tctcagaaga
      361 caaaaaaggc tttgaaactg aacaatctgg tctgggtgtc cttaggccat ccctcaccaa
      421 agagcctatt tctgtcttct gtccatgcca aataaatgat tctgagtggt ttgccttgag
      481 tccacaggag aggtatgctt ttagttcata tataagaaat gctcgttgtt ttttgtgcca
      541 tgggaaatgc aaatgtttcc ccgatggaga aatggcccat gttcttggcc aaatcattga
      601 gatctctgac tggaatgata agtgagcaaa catgtggtgt ctcccctact ccaatctacc
      661 tttgctttat cttttagatt ggcttgggca aaagaatcca gagaccctag gatttcagaa
      721 ggccagcagt ctccactgga gaagaagatc ctggtaagta atctacagtg gagctactta
      781 gaaaaaggac cagggtttaa aatccttcag cacaggtgac cctttgaggc attactctgt
      841 tccactctgt taagtattcc agatgcatcc cctttaaaaa cgggtcatct attggggatg
      901 ggcactatct actttgagaa cacccaggct tgcctgggag catggagtgt aagatgtaga
      961 gctcgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga
     1021 tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
     1081 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1141 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     1201 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     1261 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1321 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1381 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1441 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1501 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     1561 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     1621 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     1681 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1741 cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg
     1801 aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1861 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     1921 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc
     1981 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2041 cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg
     2101 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2161 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2221 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct
     2281 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2341 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2401 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     2461 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     2521 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2581 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2641 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2701 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     2761 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2821 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     2881 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2941 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     3001 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     3061 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3121 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     3181 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3241 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3301 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3361 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3421 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3481 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3541 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3601 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3661 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3721 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3781 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3841 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3901 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3961 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4021 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4081 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4141 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4201 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4261 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4321 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4381 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4441 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4501 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4561 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4621 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4681 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4741 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4801 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4861 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4921 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4981 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
