LOCUS       sequence-432577-        4906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2499m 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 4906)
  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 4906)
  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: AiP1873 - pAAV-AiE2499m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   374..503
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1291..1300
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1297..1299,1300..1302,1303..2013)
                     /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(1342..1363)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1603..1622)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1950..1971
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2106..2126)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2313..2424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2542..2682
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2542..2671
                     /label=AAV2 ITR
     rep_origin      2757..3212
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2844..2863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3054..3075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3229..3248)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3348..3370
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3408..3426)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3494..3598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3599..3667,3668..4459)
                     /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(3817..3836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4630..4906,1..312)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
       61 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      361 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
      421 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
      481 caactccatc actaggggtt cctgcggccg cacgcgttgt aagagtaata atcgaatggt
      541 tactgtatat tgcacatttt actgaacgga cctcagcagg tcgctgtctc tttcacatta
      601 cagaatcctg gagcaaaaag gggctgctgt ttagttccag gcctagctgt ctcctcagtg
      661 cactgcggga actcactaag catctattgg gctggctgca aagcacggaa atctaaattt
      721 gggtgtcagg aatatatttg ggtgtttcca gttcagggac actccagcag ctccaggtct
      781 gacccaaatt cgtcctttta agggcctgtg tagctcagtg ccatagaaca acttcatgtc
      841 cctctgtagt cccatgtcgt aatttatcag ctgtttcagg gatgcatact aagtcctgtg
      901 aagctggtac aggaacaaag aatccttgca gagcagcttg gcgatttagc tcaaatcaat
      961 tctataaata tttagcattt atgctgattg ctgccttcct accaccatat tatctttaga
     1021 ctttaataat atactgcaag attaaatgga aagtgcagcc catgatgtct gaatttccct
     1081 ttgttttatg aaaggcagta tacagctaat tttaaaagaa atagacatta ttcttaggaa
     1141 aataatatga ttttctatac tatttccttt ctaaacagaa ttttacttat ttttccggag
     1201 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
     1261 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
     1321 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1381 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
     1441 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
     1501 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1561 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1621 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1681 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1741 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     1801 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     1861 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     1921 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1981 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     2041 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2101 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2161 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2221 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2281 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2341 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2401 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2461 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2521 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2581 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2641 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2701 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     2761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2821 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2941 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     3001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3061 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     3121 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3181 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3241 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3301 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3361 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3421 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4141 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4741 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggac
//
