LOCUS       sequence-471045-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2279m 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 4840)
  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 4840)
  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: AiP1612 - pAAV-AiE2279m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4840
                     /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      852..861
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(858..860,861..863,864..1574)
                     /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(903..924)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1164..1183)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1511..1532
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1667..1687)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1874..1985
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2103..2243
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2103..2232
                     /label=AAV2 ITR
     rep_origin      2318..2773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2405..2424)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2615..2636
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2790..2809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2909..2931
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2969..2987)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3055..3159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3160..3228,3229..4020)
                     /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(3378..3397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4191..4779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4680..4699
                     /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 gcgttgtaag agtaataatc gccggcgcaa aaataactgt
      181 ccatggttcc agaatgttct cttgggtggg gttgggggga gacaaaagaa agccaaggaa
      241 cttaatgtct agagatgata attctagcag tgcgtactga ctacatttat gaaacagtat
      301 tgtgacctgg gctttgtgac tggtgctgga gacggctcac cctgatgagt taaaactcct
      361 tcatgaacca gtctcctctg ccctgtaccg tttggaaaat ggtgatgcct agagggcacc
      421 cgaatgcaga ctgtgcatgt tttggttcct gtctagacca ctcaacccaa actgaaatag
      481 ctgagctgac caaggagctg aatagggagt taaagcgggg aggggaaggg gccaggatgc
      541 ttaaaaatag tgggaaacac gttgcccaag ggctcagaga taaacagtgc agtttcaatg
      601 tctagtccta ccactaaggt catggggctt ggggaagttg ttgttttttt tttttttttc
      661 tgggtcaaag gtgcctatgg gcgagggact aggggagtgg gagggagttg gttgttggag
      721 aggtggggat aggaaaaggt attcactttc cgccggcgga gctcgggctg ggcataaaag
      781 tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag
      841 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      901 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      961 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
     1021 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct
     1081 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1141 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1201 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1261 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     1321 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     1381 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1441 tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga
     1501 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1561 acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac
     1621 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1681 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1741 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     1801 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     1861 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1921 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1981 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2041 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     2101 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2161 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     2221 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     2281 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     2341 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     2401 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2461 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2521 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2581 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2641 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     2701 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2761 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2821 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2881 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2941 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3001 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3061 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3121 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3181 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3241 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3301 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3361 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3421 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3481 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3541 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3601 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3661 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3721 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3781 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3841 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3901 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3961 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4021 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4081 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4141 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4201 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4261 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4321 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4381 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4441 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4501 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4561 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4621 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4681 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4741 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4801 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
