LOCUS       sequence-466338-        4669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2605m 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 4669)
  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 4669)
  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: AiP2031 - pAAV-AiE2605m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      229..238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(235..237,238..240,241..951)
                     /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(280..301)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(541..560)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     888..909
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1044..1064)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1251..1362
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1480..1620
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1480..1609
                     /label=AAV2 ITR
     rep_origin      1695..2150
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1782..1801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1992..2013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2167..2186)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2286..2308
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2346..2364)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2432..2536
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2537..2605,2606..3397)
                     /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(2755..2774)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3568..4156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4057..4076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4218..4347
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tcaaattatg ttccctgggg gtaagtatta taatacaccc catcttcatg atctcgctct
       61 gccagtgatt taaattttat gggcttataa tctacacact gtccttgttc ctgatataaa
      121 aacaccagaa caccggagct cgggctgggc ataaaagtca gggcagagcc atctattgct
      181 tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg
      241 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      301 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      361 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      421 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      481 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      541 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      601 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      661 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
      721 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
      781 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      841 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
      901 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac
      961 ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa
     1021 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     1081 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     1141 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     1201 ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc
     1261 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1321 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1381 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1441 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     1501 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     1561 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     1621 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     1681 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1741 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     1801 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1861 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     1921 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     1981 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     2041 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     2101 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2161 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2221 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2281 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2341 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2401 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2461 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2521 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2581 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2641 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2701 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2761 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2821 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2881 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2941 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3001 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3061 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3121 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3181 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3241 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3301 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3361 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3421 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3481 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3541 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3601 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3661 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3721 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3781 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3841 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3901 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3961 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4021 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4081 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4141 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4201 gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     4261 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     4321 tggccaactc catcactagg ggttcctgcg gccgcacgcg taacatcaca tagagaatta
     4381 gactgcagat tgtgacacaa ataaatctac ttggctttta gaaacattcc ttaaaaagaa
     4441 atattttggt accagaatgt gttcaggagg gaaagagcct gtgcccttga cacaggaagt
     4501 agataacacc tgtcagaaga atcgaaacca cctgcttggg tgcatctggt aataagtttg
     4561 cagctggtgc tacactctac cactatggac cataactcca ggcaatagag aaggttcata
     4621 gtggacaaaa cccttcacaa tctggaagat cccagctatg ggattcctg
//
