LOCUS       sequence-432551-        4860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2407m 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 4860)
  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 4860)
  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: AiP1708 - pAAV-AiE2407m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..48
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   190..319
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1067..1069,1070..1072,1073..1783)
                     /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(1112..1133)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1373..1392)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1720..1741
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1876..1896)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2083..2194
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2312..2452
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2312..2441
                     /label=AAV2 ITR
     rep_origin      2527..2982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2614..2633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2824..2845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2999..3018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3118..3140
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3178..3196)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3264..3368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3369..3437,3438..4229)
                     /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(3587..3606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4400..4860,1..128)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
       61 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      121 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      181 ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg
      241 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
      301 tccatcacta ggggttcctg cggccgcacg cgttgtaaga gtaataatcg acccgcctgg
      361 cattgctgct gaacattaag acacaaatgg aaatgaaaca cattgacaaa taccatccgt
      421 ggcatggggt agagccatcc aaggtgtgcc gtgtgacaag aagatgagga gtcctttggg
      481 atgagtcacc tggccagggt caaatgagag gcagtacaga gactagaaac cagctctccc
      541 caatctgact ctcctgagaa ggggctctgt ccccacataa gtcactcatc tcccactctg
      601 attctccctg ttcaaaacat ctctgtattc ataggtccaa cggacaccca cacctccttc
      661 ccactttaag ctcctaaatg caatcatgct acgtactcag acctatatta gccatgcaag
      721 aaggatgcag tgtgtatcca agaacaaaca cttcattaag ccctagattt aattaactta
      781 aatcattttc catcccagct cccatgtccc acttgaaata gagagccatt ttaagtgtag
      841 tccctaactc ttaactagaa gttctgattt tagtggtttt gaatcagact cgacattaat
      901 agctcactgt cttcatctgc ccccgaggta gcttcctccc ccatgtgatt aatttcattt
      961 ttctccggag ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg
     1021 cttctgggat ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg
     1081 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1141 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1201 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1261 gggctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1321 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1381 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1441 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1501 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     1561 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     1621 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     1681 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1741 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc
     1801 gcggccgcga attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac
     1861 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1921 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1981 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2041 gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc
     2101 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2161 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2221 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2281 atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     2341 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2401 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     2461 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     2521 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2581 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     2641 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2701 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     2761 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2821 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     2881 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     2941 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3001 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3061 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3121 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3181 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3241 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3301 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3361 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3421 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3481 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3541 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3601 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3661 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3721 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3781 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3841 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3901 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3961 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4021 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4081 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4141 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4201 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4261 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4321 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4381 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4441 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4501 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4561 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4621 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4681 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4741 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4801 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
//
