LOCUS       sequence-466212-        5763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2053m 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 5763)
  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 5763)
  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: AiP1213 - pAAV-AiE2053m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(276..295)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     395..417
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(455..473)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        541..645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(646..714,715..1506)
                     /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(864..883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1677..2265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2166..2185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2327..2456
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2887..2896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2893..2898,2899..2919,2920..4191)
                     /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    4216..4804
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4269..4289)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4687..4698)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4836..5312
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5035..5054)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5352..5492
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5352..5481
                     /label=AAV2 ITR
     rep_origin      join(5567..5763,1..259)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5654..5673)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
       61 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      121 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
      181 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      241 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
      301 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
      361 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
      421 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
      481 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
      541 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      601 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      661 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      721 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      781 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      841 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      901 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      961 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1021 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1081 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1141 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1201 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1261 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1321 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1381 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1441 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1501 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1561 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1621 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1681 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1741 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1801 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1861 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1921 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1981 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2041 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2101 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2161 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2221 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2281 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
     2341 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc
     2401 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
     2461 ccgcacgcgt agaggttatt tggcttgctc aggatcaaca aattttaaat gtcagcgcag
     2521 attcagagcc agagcatatg acttcaaggc gggtactcat tgcaccacta tctggcggtg
     2581 gccttttctt tcagcccaca gtagaactag tctcaaacta aagggcactt accattttgc
     2641 tggcataact gagcagctaa atgtgttgac acattccttc ttgtgtcttt atagctgttt
     2701 cctgactgcc ttgtctttac agctcctaaa ctccttccat atataaccaa tgtcagagac
     2761 agcagttggt cattgtcatt acacttaatg atttggtgga gaatctacca actgccctta
     2821 aggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
     2881 ggcgtggcca ccatggctcc taagaagaag aggaaggtga tgagccagtt cgacatcctg
     2941 tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg agagattcga gaggcccagc
     3001 ggcgagaaga tcgccagctg tgccgccgag ctgacctacc tgtgctggat gatcacccac
     3061 aacggcaccg ccatcaagag ggccaccttc atgagctaca acaccatcat cagcaacagc
     3121 ctgagcttcg acatcgtgaa caagagcctg cagttcaagt acaagaccca gaaggccacc
     3181 atcctggagg ccagcctgaa gaagctgatc cccgcctggg agttcaccat catcccttac
     3241 aacggccaga agcaccagag cgacatcacc gacatcgtgt ccagcctgca gctgcagttc
     3301 gagagcagcg aggaggccga caagggcaac agccacagca agaagatgct gaaggccctg
     3361 ctgtccgagg gcgagagcat ctgggagatc accgagaaga tcctgaacag cttcgagtac
     3421 accagcaggt tcaccaagac caagaccctg taccagttcc tgttcctggc cacattcatc
     3481 aactgcggca ggttcagcga catcaagaac gtggacccca agagcttcaa gctggtgcag
     3541 aacaagtacc tgggcgtgat cattcagtgc ctggtgaccg agaccaagac aagcgtgtcc
     3601 aggcacatct actttttcag cgccagaggc aggatcgacc ccctggtgta cctggacgag
     3661 ttcctgagga acagcgagcc cgtgctgaag agagtgaaca ggaccggcaa cagcagcagc
     3721 aacaagcagg agtaccagct gctgaaggac aacctggtgc gcagctacaa caaggccctg
     3781 aagaagaacg ccccctaccc catcttcgct atcaagaacg gccctaagag ccacatcggc
     3841 aggcacctga tgaccagctt tctgagcatg aagggcctga ccgagctgac aaacgtggtg
     3901 ggcaactgga gcgacaagag ggcctccgcc gtggccagga ccacctacac ccaccagatc
     3961 accgccatcc ccgaccacta cttcgccctg gtgtccaggt actacgccta cgaccccatc
     4021 agcaaggaga tgatcgccct gaaggacgag accaacccca tcgaggagtg gcagcacatc
     4081 gagcagctga agggcagcgc cgagggcagc atcagatacc ccgcctggaa cggcatcatc
     4141 agccaggagg tgctggacta cctgagcagc tacatcaaca ggcggatctg agaattcgat
     4201 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4261 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4321 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4381 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4441 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4501 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4561 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     4621 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     4681 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4741 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4801 ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     4861 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     4921 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     4981 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     5041 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     5101 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     5161 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     5221 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     5281 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac
     5341 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     5401 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     5461 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     5521 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     5581 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5641 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5701 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5761 ctc
//
