LOCUS       sequence-462013-        4459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2473m 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 4459)
  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 4459)
  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: AiP1934 - pAAV-AiE2473m-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4459
                     /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      471..480
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(477..479,480..482,483..1193)
                     /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(522..543)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(783..802)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1130..1151
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1286..1306)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1493..1604
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1722..1862
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1722..1851
                     /label=AAV2 ITR
     rep_origin      1937..2392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2024..2043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2234..2255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2409..2428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2528..2550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2588..2606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2674..2778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2779..2847,2848..3639)
                     /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(2997..3016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3810..4398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4299..4318
                     /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 gcgttgtaag agtaataatc gtggtgccat ggatgacctc
      181 agtttccccg agtccctatg gcttatgaat caaattaatc acttccccac ctacgccaca
      241 gctcagggtc ccctgccagc agattccaga cgcactagga ttctggtcca aacaggccct
      301 gagcagtgta accctgagca ctgattggct cttcttcgct cctcgtgact cctcccctca
      361 tccctggttt tgcctaagag cctggtgcag cctcaccagc tctgggtagg tgctctggtt
      421 ctcaccaagg aagcttcctt gggtgcacca ggcttaatgc gctaccggtc gccaccatgg
      481 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      541 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      601 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      661 tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
      721 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      781 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      841 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
      901 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
      961 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     1021 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1081 tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1141 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg
     1201 acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg
     1261 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1321 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1381 aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1441 caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct
     1501 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1561 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1621 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1681 aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     1741 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     1801 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     1861 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     1921 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1981 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     2041 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2101 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2161 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2221 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     2281 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     2341 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     2401 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     2461 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2521 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     2581 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2641 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2701 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2761 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2821 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2881 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2941 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3001 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3061 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3121 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3181 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3241 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3301 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3361 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3421 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3481 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3541 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3601 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3661 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3721 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3781 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3841 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3901 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3961 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4021 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4081 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4141 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4201 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4261 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4321 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4381 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4441 tggccttttg ctcacatgt
//
