LOCUS       sequence-462005-        4749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2539m 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 4749)
  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 4749)
  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: AiP1859 - pAAV-AiE2539m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4749
                     /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      761..770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(767..769,770..772,773..1483)
                     /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(812..833)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1073..1092)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1420..1441
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1576..1596)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1783..1894
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2012..2152
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2012..2141
                     /label=AAV2 ITR
     rep_origin      2227..2682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2314..2333)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2524..2545
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2699..2718)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2818..2840
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2878..2896)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2964..3068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3069..3137,3138..3929)
                     /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(3287..3306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4100..4688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4589..4608
                     /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 gtccagagct gtgcccatca
      181 caaagtggta ctgcacgtct ctttgacgca cttgtggccc ggtgacgttc tgcagtcgct
      241 cgtcggtgca cagaagccag ggagcagggg aactcccggg catcgatgct ctgtgtccag
      301 ctcaaaaaac tctgataaaa atagctggtg agtaagaacc cgggtgggaa gtgggccagc
      361 agccagagag ggacacagca ggagagcccg cagctctgca tggaaggatc agatcgtcag
      421 accactgaca cctccatctt gcagcctgtt gtggtccctc gacctctgat tggtcactgg
      481 aaggggacag tgggctcggg gtgcactgca tgccccttca ggtctgagcg gagcatacct
      541 tttcagatct ggctccctag agggactttc tctttgtctc aacttcccca gaatccaaca
      601 ctctcccagg tcacaaggac attcttgata gtatttccca ggaaatctgt ggctagtccc
      661 tctcccggag ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg
      721 cttctgggat ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg
      781 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      841 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      901 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      961 gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1021 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1081 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1141 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1201 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     1261 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     1321 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     1381 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1441 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc
     1501 gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac
     1561 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1621 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1681 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     1741 gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc
     1801 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1861 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1921 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1981 atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     2041 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2101 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     2161 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     2221 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2281 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     2341 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2401 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     2461 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2521 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     2581 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     2641 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     2701 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     2761 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2821 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     2881 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     2941 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3001 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3061 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3121 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3181 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3241 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3301 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3361 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3421 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3481 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3541 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3601 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3661 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3721 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3781 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3841 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3901 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3961 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4021 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4081 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4141 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4201 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4261 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4321 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4381 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4441 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4501 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4561 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4621 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4681 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4741 ctcacatgt
//
