LOCUS       sequence-453078-        5800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2050m 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 5800)
  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 5800)
  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: AiP1215 - pAAV-AiE2050m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5800
                     /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      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(604..609,610..630,631..1902)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    1927..2515
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1980..2000)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2398..2409)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2547..3023
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2746..2765)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3063..3203
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3063..3192
                     /label=AAV2 ITR
     rep_origin      3278..3733
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3365..3384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3575..3596
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3750..3769)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3869..3891
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3929..3947)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4015..4119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4120..4188,4189..4980)
                     /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(4338..4357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5151..5739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5640..5659
                     /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 gcgtaaagac cagcattgcc attgggagca ggcacagctc
      181 ctctgatgga cacacagctc ctctcctgag caactgcact atctgttcct cctgggatgc
      241 ctgcctctca cccacatcct acccactgtt ggccctcatt ttcacactca tttgtcatgg
      301 cttcttggga aatgtgtgct ataaaaggac cgtgtgcaac gtgaaggtca ctgtactggt
      361 gctgtgcctt gggccctcta gaaaaaagtg ttctgaagac aagtgcagtg gagaaatgcc
      421 aggaggccta cagaggtatt tcactctcca tctgcatttg cggtgcaatg acacaggcag
      481 ccatgctgga cggggcctcc cggaggcaag aggagaggct ctgctgactt aaggagctcg
      541 ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc tggcgtggcc
      601 accatggctc ctaagaagaa gaggaaggtg atgagccagt tcgacatcct gtgcaagacc
      661 ccccccaagg tgctggtgcg gcagttcgtg gagagattcg agaggcccag cggcgagaag
      721 atcgccagct gtgccgccga gctgacctac ctgtgctgga tgatcaccca caacggcacc
      781 gccatcaaga gggccacctt catgagctac aacaccatca tcagcaacag cctgagcttc
      841 gacatcgtga acaagagcct gcagttcaag tacaagaccc agaaggccac catcctggag
      901 gccagcctga agaagctgat ccccgcctgg gagttcacca tcatccctta caacggccag
      961 aagcaccaga gcgacatcac cgacatcgtg tccagcctgc agctgcagtt cgagagcagc
     1021 gaggaggccg acaagggcaa cagccacagc aagaagatgc tgaaggccct gctgtccgag
     1081 ggcgagagca tctgggagat caccgagaag atcctgaaca gcttcgagta caccagcagg
     1141 ttcaccaaga ccaagaccct gtaccagttc ctgttcctgg ccacattcat caactgcggc
     1201 aggttcagcg acatcaagaa cgtggacccc aagagcttca agctggtgca gaacaagtac
     1261 ctgggcgtga tcattcagtg cctggtgacc gagaccaaga caagcgtgtc caggcacatc
     1321 tactttttca gcgccagagg caggatcgac cccctggtgt acctggacga gttcctgagg
     1381 aacagcgagc ccgtgctgaa gagagtgaac aggaccggca acagcagcag caacaagcag
     1441 gagtaccagc tgctgaagga caacctggtg cgcagctaca acaaggccct gaagaagaac
     1501 gccccctacc ccatcttcgc tatcaagaac ggccctaaga gccacatcgg caggcacctg
     1561 atgaccagct ttctgagcat gaagggcctg accgagctga caaacgtggt gggcaactgg
     1621 agcgacaaga gggcctccgc cgtggccagg accacctaca cccaccagat caccgccatc
     1681 cccgaccact acttcgccct ggtgtccagg tactacgcct acgaccccat cagcaaggag
     1741 atgatcgccc tgaaggacga gaccaacccc atcgaggagt ggcagcacat cgagcagctg
     1801 aagggcagcg ccgagggcag catcagatac cccgcctgga acggcatcat cagccaggag
     1861 gtgctggact acctgagcag ctacatcaac aggcggatct gagaattcga tatcaagctt
     1921 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1981 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2041 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2101 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2161 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2221 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2281 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     2341 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2401 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2461 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     2521 taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc
     2581 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     2641 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     2701 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     2761 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     2821 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     2881 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     2941 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     3001 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc
     3061 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3121 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3181 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3241 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3301 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3361 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3421 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3481 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3541 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3601 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     3661 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3721 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3781 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3841 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3901 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3961 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4021 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4081 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4141 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4201 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4261 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4321 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4381 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4441 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4501 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4561 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4621 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4681 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4741 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4801 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4861 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4921 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4981 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5041 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5101 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5161 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5221 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5281 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5341 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5401 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5461 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5521 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5581 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5641 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5701 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5761 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
