LOCUS       sequence-466191-        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"
     rep_origin      200..788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     689..708
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   850..979
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1447..1456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1453..1458,1459..1479,1480..2751)
                     /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    2776..3364
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2829..2849)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3247..3258)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3396..3872
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3595..3614)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3912..4052
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3912..4041
                     /label=AAV2 ITR
     rep_origin      4127..4582
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4214..4233)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4424..4445
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4599..4618)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4718..4740
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4778..4796)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4864..4968
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4969..5037,5038..5800,1..29)
                     /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(5187..5206)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
       61 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
      121 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
      181 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      241 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      301 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      361 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      421 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      481 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      541 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      601 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      661 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      721 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      781 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      841 ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg
      901 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
      961 tccatcacta ggggttcctg cggccgcacg cgtaaagacc agcattgcca ttgggagcag
     1021 gcacagctcc tctgatggac acacagctcc tctcctgagc aactgcacta tctgttcctc
     1081 ctgggatgcc tgcctctcac ccacatccta cccactgttg gccctcattt tcacactcat
     1141 ttgtcatggc ttcttgggaa atgtgtgcta taaaaggacc gtgtgcaacg tgaaggtcac
     1201 tgtactggtg ctgtgccttg ggccctctag aaaaaagtgt tctgaagaca agtgcagtgg
     1261 agaaatgcca ggaggcctac agaggtattt cactctccat ctgcatttgc ggtgcaatga
     1321 cacaggcagc catgctggac ggggcctccc ggaggcaaga ggagaggctc tgctgactta
     1381 aggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
     1441 ggcgtggcca ccatggctcc taagaagaag aggaaggtga tgagccagtt cgacatcctg
     1501 tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg agagattcga gaggcccagc
     1561 ggcgagaaga tcgccagctg tgccgccgag ctgacctacc tgtgctggat gatcacccac
     1621 aacggcaccg ccatcaagag ggccaccttc atgagctaca acaccatcat cagcaacagc
     1681 ctgagcttcg acatcgtgaa caagagcctg cagttcaagt acaagaccca gaaggccacc
     1741 atcctggagg ccagcctgaa gaagctgatc cccgcctggg agttcaccat catcccttac
     1801 aacggccaga agcaccagag cgacatcacc gacatcgtgt ccagcctgca gctgcagttc
     1861 gagagcagcg aggaggccga caagggcaac agccacagca agaagatgct gaaggccctg
     1921 ctgtccgagg gcgagagcat ctgggagatc accgagaaga tcctgaacag cttcgagtac
     1981 accagcaggt tcaccaagac caagaccctg taccagttcc tgttcctggc cacattcatc
     2041 aactgcggca ggttcagcga catcaagaac gtggacccca agagcttcaa gctggtgcag
     2101 aacaagtacc tgggcgtgat cattcagtgc ctggtgaccg agaccaagac aagcgtgtcc
     2161 aggcacatct actttttcag cgccagaggc aggatcgacc ccctggtgta cctggacgag
     2221 ttcctgagga acagcgagcc cgtgctgaag agagtgaaca ggaccggcaa cagcagcagc
     2281 aacaagcagg agtaccagct gctgaaggac aacctggtgc gcagctacaa caaggccctg
     2341 aagaagaacg ccccctaccc catcttcgct atcaagaacg gccctaagag ccacatcggc
     2401 aggcacctga tgaccagctt tctgagcatg aagggcctga ccgagctgac aaacgtggtg
     2461 ggcaactgga gcgacaagag ggcctccgcc gtggccagga ccacctacac ccaccagatc
     2521 accgccatcc ccgaccacta cttcgccctg gtgtccaggt actacgccta cgaccccatc
     2581 agcaaggaga tgatcgccct gaaggacgag accaacccca tcgaggagtg gcagcacatc
     2641 gagcagctga agggcagcgc cgagggcagc atcagatacc ccgcctggaa cggcatcatc
     2701 agccaggagg tgctggacta cctgagcagc tacatcaaca ggcggatctg agaattcgat
     2761 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2821 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2881 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2941 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3001 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3061 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3121 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3181 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3241 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3301 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3361 ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     3421 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     3481 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     3541 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     3601 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     3661 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     3721 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     3781 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     3841 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac
     3901 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3961 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4021 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     4081 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     4141 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4201 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4261 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4321 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4381 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4441 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     4501 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4561 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     4621 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4681 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4741 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4801 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4861 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4921 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4981 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5041 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5101 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5161 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5221 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5281 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5341 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5401 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5461 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5521 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5581 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5641 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5701 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5761 caggcaacta tggatgaacg aaatagacag atcgctgaga
//
