LOCUS       sequence-466285-        4608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2345m 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 4608)
  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 4608)
  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: AiP1662 - pAAV-AiE2345m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     208..230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(268..286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        354..458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(459..527,528..1319)
                     /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(677..696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1490..2078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1979..1998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2140..2269
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2759..2768
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2765..2767,2768..2770,2771..3481)
                     /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(2810..2831)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3071..3090)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3418..3439
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3574..3594)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3781..3892
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4010..4150
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4010..4139
                     /label=AAV2 ITR
     rep_origin      join(4225..4608,1..72)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4312..4331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4522..4543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
       61 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      121 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
      181 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
      241 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
      301 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
      361 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      421 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      481 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      541 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
      601 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
      661 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
      721 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
      781 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
      841 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
      901 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
      961 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1021 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1081 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1141 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1201 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1261 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1321 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1381 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1441 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1501 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1561 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1621 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1681 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1741 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1801 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1861 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1921 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1981 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2041 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2101 tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc
     2161 gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     2221 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg
     2281 cgttgtaaga gtaataatcg gagcacaacc acagaatgac acagaagtta gtgttctggt
     2341 gtctcagact tgaaactgcc acggctctag aggtgatggt gcctgggaga atgcagccaa
     2401 taaatgaggc cgactaaatt gtgtgccttt attcagatcc tcagacagtt taagaactct
     2461 gagggttaag aaagctcgca ggggtaaagt tttcatcagt tcaagaatta cacacccatc
     2521 aggacaagac acacttgtca aaatcatgtt ttattcatag aggcatttaa acatttaact
     2581 taatctgtga tagttgggag gagcacaaaa acacattctc agagcaattc catattttga
     2641 agacattgat gtcacaagac tctcggagct cgggctgggc ataaaagtca gggcagagcc
     2701 atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc
     2761 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2821 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2881 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
     2941 caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat
     3001 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3061 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3121 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3181 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     3241 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     3301 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3361 ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     3421 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3481 gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa
     3541 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3601 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3661 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     3721 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga
     3781 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3841 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3901 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3961 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     4021 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4081 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4141 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4201 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4261 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     4321 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4381 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4441 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4501 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     4561 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtct
//
