LOCUS       sequence-462019-        4677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2597m 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 4677)
  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 4677)
  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: AiP2022 - pAAV-AiE2597m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4677
                     /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      689..698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(695..697,698..700,701..1411)
                     /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(740..761)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1001..1020)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1348..1369
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1504..1524)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1711..1822
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1940..2080
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1940..2069
                     /label=AAV2 ITR
     rep_origin      2155..2610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2242..2261)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2452..2473
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2627..2646)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2746..2768
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2806..2824)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2892..2996
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2997..3065,3066..3857)
                     /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(3215..3234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4028..4616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4517..4536
                     /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 attctctata aaactgtcca cagaggcaca actgccagcc
      181 cagggtgtgc gcatgtgcgg gttttgctta agtttcttgt ttctgacttt gcatctgcgc
      241 aacactgaga agtgtcaagt cttctcccac cattagatga aagttggaat tttgagcttt
      301 tggaaggacc gtctcagttt caacactgaa atattcatcg ttgaaaaagc tctctcttag
      361 gaggggaaaa ataagatgta aattggtttc atttaaatct cactacttgc atcactcagg
      421 cctgctaagc tgatagagcc aggctggaaa gggctgtctc cctcctgcgt catctggagc
      481 agggcagagg aaccccagct gcttctgtct ggggcaggtt gccagctgag ggcccaagga
      541 gagctccagg ctcccagggc tactactgct attgtggaaa gtgaaagact tatcggagct
      601 cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc
      661 agatctttcg aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt
      721 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      781 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
      841 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt
      901 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      961 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1021 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1081 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1141 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1201 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     1261 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
     1321 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1381 gatcactctc ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat
     1441 tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1501 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1561 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     1621 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     1681 tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg
     1741 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1801 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1861 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac
     1921 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     1981 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     2041 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     2101 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     2161 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2221 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2281 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2341 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     2401 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2461 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     2521 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2581 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     2641 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2701 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2761 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     2821 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2881 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2941 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3001 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3061 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3121 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3181 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3241 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3301 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3361 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3421 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3481 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3541 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3601 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3661 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3721 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3781 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3841 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3901 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3961 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4021 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4081 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4141 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4201 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4261 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4321 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4381 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4441 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4501 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4561 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4621 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
