LOCUS       sequence-466256-        5035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2171m 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 5035)
  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 5035)
  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: AiP1400 - pAAV-AiE2171m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     425..444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   586..715
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1632..1641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1638..1640,1641..1643,1644..2354)
                     /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(1683..1704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1944..1963)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2291..2312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2447..2467)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2654..2765
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2883..3023
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2883..3012
                     /label=AAV2 ITR
     rep_origin      3098..3553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3185..3204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3395..3416
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3570..3589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3689..3711
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3749..3767)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3835..3939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3940..4008,4009..4800)
                     /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(4158..4177)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4971..5035,1..524)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
       61 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      121 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      181 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      241 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      301 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      361 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      421 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      481 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      541 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc
      601 gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca
      661 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
      721 cgcacgcgta cacacgggct gtcggtgtgt gcatctcatg ggaacacttg ccctgccagc
      781 cctggggtag gtatatccct tgggtggtgg taactgagct gtggcctggg cagcttctag
      841 tggcccagag aaacactgtt ttggctagaa acaacttcct tccagacgaa gagggcctga
      901 tgggaaaggg ccttgactca gccaagttgg actgtctcac tgatccaccc atcatcaagg
      961 gggcctcctt gtccccagct tcactaagaa taaactttct ggtcctttct cctgctgcta
     1021 tgtgtaatcc cagtcccagc agctggcaaa ggggcccatg cctccctcaa caacctccag
     1081 gcctgtttat tcccttggac agaagaaaga aaaaaaaacc acaatctctg tcaggactaa
     1141 acaatgagga acagaaagcc cctgcgtaat gcagaacaaa aggtcactcc tgcaaaggga
     1201 ggtacccaaa gccccaaaac atagtcttgg gggctgggcc tcagggccac tacagggact
     1261 gcagagggat gggggtggga atgggggcac tagagagggt tcacaagtcc ggagattccc
     1321 aaggctgggg aggactagtc ttttacttta aattagagag acacagattc acccccaccc
     1381 ccagtgcatg gagatcagac actgagaaat gaggaaatga atccaacttc acttgatgct
     1441 gtcagcatgt ttgtgtgagc cagttgctca gaagataaag cctccagcag ggagtagtgg
     1501 agtggcatat gcctttaatc ccagagctaa gttggtggga gctcgggctg ggcataaaag
     1561 tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag
     1621 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1681 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1741 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
     1801 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct
     1861 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1921 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1981 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2041 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     2101 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     2161 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2221 tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga
     2281 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2341 acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac
     2401 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2461 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2521 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     2581 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc
     2641 gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2701 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2761 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2821 aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc
     2881 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2941 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3001 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3061 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3121 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3181 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3241 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3301 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3361 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3421 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     3481 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3541 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3601 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3661 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3721 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3781 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3901 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3961 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4021 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4081 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4141 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4201 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4261 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4321 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4381 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4441 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4501 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4561 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4621 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4681 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4741 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4801 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcgg
//
