LOCUS       sequence-466221-        5939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2068m 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 5939)
  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 5939)
  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: AiP1240 - pAAV-AiE2068m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   89..229
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   89..218
                     /label=AAV2 ITR
     rep_origin      304..759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(391..410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     601..622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(776..795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     895..917
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(955..973)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1041..1145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1146..1214,1215..2006)
                     /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(1364..1383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2177..2765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2666..2685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2827..2956
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3563..3572
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3569..3574,3575..3595,3596..4867)
                     /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    4892..5480
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4945..4965)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5363..5374)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(5512..5939,1..49)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5711..5730)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
       61 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
      121 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
      181 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
      241 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
      301 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      361 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
      421 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      481 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
      541 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      601 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
      661 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
      721 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
      781 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
      841 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
      901 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
      961 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     1021 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1081 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1141 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1201 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1261 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1321 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1381 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1441 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1501 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1561 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1621 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1681 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1741 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1801 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1861 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1921 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1981 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2041 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2101 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2161 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2221 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2281 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2341 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2401 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2461 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2521 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2581 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2641 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2701 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2761 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2821 acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg
     2881 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     2941 atcactaggg gttcctgcgg ccgcacgcgt ccaggaagaa acaaagaagc agctttagtc
     3001 aacaagatcc tttcagaaga ccaaatgcac aaccatccct caggcctcta caagctgctc
     3061 agtgtgcagg gctctcgcct cctcagacca cgctcagggg ttgctaacgg cacctggtat
     3121 tattgagaac ttagcaacgt cagatcaaat gcttctgttc atggtctgac ccagtcctta
     3181 ctatgagttc agatttatga ctctcaattt actaaaaaag aaactgatga tttgagagga
     3241 gaaatgaccc agctaaggcc atgttaggaa tgtcttgatg agccaagact tcatccagac
     3301 ctgattctgg ggcctgtgct gtcatccaat gtgcctggcc actaaagaac ttcagttcct
     3361 tgtggaatgg ggccagatgc cccaccccca gatagttcag caccttgtat gctccttgag
     3421 actctgtaaa aaaatttttt aagccaacaa atgactgaaa aactacttga acaaaagcca
     3481 agcatgtttg ttcagaccga gctcgggctg ggcataaaag tcagggcaga gccatctatt
     3541 gcttacattt gcttctggcg tggccaccat ggctcctaag aagaagagga aggtgatgag
     3601 ccagttcgac atcctgtgca agaccccccc caaggtgctg gtgcggcagt tcgtggagag
     3661 attcgagagg cccagcggcg agaagatcgc cagctgtgcc gccgagctga cctacctgtg
     3721 ctggatgatc acccacaacg gcaccgccat caagagggcc accttcatga gctacaacac
     3781 catcatcagc aacagcctga gcttcgacat cgtgaacaag agcctgcagt tcaagtacaa
     3841 gacccagaag gccaccatcc tggaggccag cctgaagaag ctgatccccg cctgggagtt
     3901 caccatcatc ccttacaacg gccagaagca ccagagcgac atcaccgaca tcgtgtccag
     3961 cctgcagctg cagttcgaga gcagcgagga ggccgacaag ggcaacagcc acagcaagaa
     4021 gatgctgaag gccctgctgt ccgagggcga gagcatctgg gagatcaccg agaagatcct
     4081 gaacagcttc gagtacacca gcaggttcac caagaccaag accctgtacc agttcctgtt
     4141 cctggccaca ttcatcaact gcggcaggtt cagcgacatc aagaacgtgg accccaagag
     4201 cttcaagctg gtgcagaaca agtacctggg cgtgatcatt cagtgcctgg tgaccgagac
     4261 caagacaagc gtgtccaggc acatctactt tttcagcgcc agaggcagga tcgaccccct
     4321 ggtgtacctg gacgagttcc tgaggaacag cgagcccgtg ctgaagagag tgaacaggac
     4381 cggcaacagc agcagcaaca agcaggagta ccagctgctg aaggacaacc tggtgcgcag
     4441 ctacaacaag gccctgaaga agaacgcccc ctaccccatc ttcgctatca agaacggccc
     4501 taagagccac atcggcaggc acctgatgac cagctttctg agcatgaagg gcctgaccga
     4561 gctgacaaac gtggtgggca actggagcga caagagggcc tccgccgtgg ccaggaccac
     4621 ctacacccac cagatcaccg ccatccccga ccactacttc gccctggtgt ccaggtacta
     4681 cgcctacgac cccatcagca aggagatgat cgccctgaag gacgagacca accccatcga
     4741 ggagtggcag cacatcgagc agctgaaggg cagcgccgag ggcagcatca gataccccgc
     4801 ctggaacggc atcatcagcc aggaggtgct ggactacctg agcagctaca tcaacaggcg
     4861 gatctgagaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     4921 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     4981 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5041 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5101 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5161 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5221 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5281 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     5341 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5401 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5461 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     5521 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     5581 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     5641 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     5701 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     5761 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     5821 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     5881 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctcc
//
