LOCUS       sequence-466352-        4984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2414m 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 4984)
  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 4984)
  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: AiP20022 - pAAV-AiE2414m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      514..523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(520..522,523..525,526..1236)
                     /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(565..586)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(826..845)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1173..1194
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1329..1349)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1536..1647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1765..1905
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1765..1894
                     /label=AAV2 ITR
     rep_origin      1980..2435
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2067..2086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2277..2298
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2452..2471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2571..2593
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2631..2649)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2717..2821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2822..2890,2891..3682)
                     /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(3040..3059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3853..4441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4342..4361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4503..4632
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 atgattgaac ttcacttcca tctgacaagg ctgattcact ggccaagccc tgactggggt
       61 gtgtgtgtga ggggtggatg gtggtggtgc ggggagatgg gtagagactg ggcggggcag
      121 ggcggggggt ggcagactct acttatgcca atttagtatg gtcatagtta gaaagaaatg
      181 ggagctggtt acctagaata gtatcttccc cagttctctg agattctttt atgcccaggt
      241 gcccctaggc ttggaagcag gacaagatcc tcaaaggatc tgttgaaagc tggctggcct
      301 tcgatcaatg atctgtgtga gctttcaagg gggaagcagg gagaggaaag caggacagac
      361 aggcactgct ctccaggaga ggcaaggtgt ttgcacttga gggagagccc tccgccggcg
      421 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
      481 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
      541 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      601 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
      661 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
      721 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      781 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      841 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      901 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      961 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1021 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1081 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1141 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1201 gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccgcggccg
     1261 cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1321 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1381 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     1441 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     1501 ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc
     1561 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1621 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1681 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat
     1741 ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     1801 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     1861 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     1921 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     1981 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2041 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2101 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2161 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2221 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2281 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     2341 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2401 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     2461 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2521 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2581 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     2641 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2701 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2761 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2821 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2881 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2941 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3001 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3061 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3121 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3181 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3241 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3301 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3361 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3421 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3481 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3541 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3601 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3661 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3721 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3781 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3841 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3901 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3961 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4021 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4081 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4141 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4201 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4261 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4321 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4381 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4441 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4501 gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct
     4561 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     4621 ctaggggttc ctgcggccgc acgcgtgaaa taggctgtcc gtagtgttta ggcttttccc
     4681 aaccaagcaa gatttcaggg atgattctta aagccacatc ttgcttcatg aaacatcaga
     4741 gagaaatctg gtcaggaaac tccagaaaat aatttctcag ccatctgtct tcccctgaat
     4801 ggcagccaga gatcgctcac caagtcatgt gtggcagctc agttggaaag agacccaagc
     4861 caaggtgcca gggtagctgt gctctgtttg gtgatggcag ctgctggcag gctgctcctg
     4921 ggtcaggggg aacagatgac ttaaggacaa gaaaatgggc tccccaaacc ttcccagtta
     4981 ttca
//
