LOCUS       sequence-466228-        7508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2104m 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 7508)
  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 7508)
  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: AiP1253 - pAAV-AiE2104m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..7508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2268..2277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2274..2279,2280..2300,2301..3572)
                     /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    3597..4185
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3650..3670)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4068..4079)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4217..4693
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4416..4435)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4733..4873
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4733..4862
                     /label=AAV2 ITR
     rep_origin      4948..5403
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5035..5054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5245..5266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5420..5439)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5539..5561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5599..5617)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5685..5789
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5790..5858,5859..6650)
                     /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(6008..6027)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6821..7409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7310..7329
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7471..7508,1..92)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
       61 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtccca gaaatcaaag
      121 acatagaaag ataattttct gtaaacaatg tttcttattg ttttgggcca tattccaatt
      181 tccaaatgta gatctattga acaaagtttc acatcaattc aaatgtagta ttgaaatgta
      241 ttttcagtgg cgaaaagtga gatatcatct tacaatgagc atcatgttca ttcctagaaa
      301 tatctcagtg ctgcagatat tgagttcata cttctcagat tcaaaattac atacctttta
      361 tgaaaatgag aactaactat tcaaacacta tcccatctga ggtcacatct tggacttgag
      421 tcccagaagc atctattctg gaatctggct gattctacta tttgttaata gggagactaa
      481 aatagcaaaa ataagccata ggaaaagaaa ataaaggaaa tactgtgtca gcccatctaa
      541 atgtattttc tgtatcatcc cttgtttctg aaagctgagg catacagact caggaatcaa
      601 taatcaacat gaattgcctt tattagacag aagtctgcac tatgcagtga gagcaatttt
      661 cctgatgaag gttgtgtaca tggagctata ttgttcccac agggtaaagt gatagggaag
      721 gtgcctactt tgttttaaat gactaaagtc aaaatagagc aaacataacc cagtagtttg
      781 cagacagtga ataagagagc tgtttcttgt ggtccagaga aaaaagatgc tgtttgtata
      841 tgctcagcct agggagtgtc actattagga ggtatagcct tggtggaata ggtgtgtcat
      901 tgcaggtatg ggctttaaga ccctcgtcct agctatctgg aagccagtct tccattagcg
      961 gccttcagat gaagatgtag aaatctcagt tctccctgca tcgtgcctgc ctagatgctg
     1021 ccaagctcct atgttgatga taatcaaata agcctctgaa cctgtaagcc agccccaatt
     1081 aaatgctgtc ctttttaaga cttgacttgg tcatggtgtc tgttcacagg agtaaagccc
     1141 aaactaagac agaagatggt accagagtct ggggtattgt tgtgataggt ctgaccatgc
     1201 ttttgtttgg aagaatatgg attggggact ttggatttgg aaagcagtgg aatcctttaa
     1261 gtagggcata atgggctatc ctagtagtaa tatggaagac tttctcgctg agtatgattt
     1321 gaacactgca gatctagtct aagtggtttc agtggagaag gattccggta tgtggtgtag
     1381 aaattatttt tgcaatattt tggtgaagaa tgtgactact ttttgccatt ggccaaagag
     1441 tcttccagag gtgagggtga agagatttag attaattgct ttgaaaaaga aagtctcaaa
     1501 aaagcctagc agagactttg ttgtctgatt aagtctcatg aagagtgttt tgaacaagta
     1561 gaccaagctt agaagtgaaa gatataaaat atatagttgg agtactaaaa gaatgccagg
     1621 aactggagct ggaccctgtg ttgaaagata ttaaattgaa ttaagggagc ggtgagtttg
     1681 gggcaagatt cccacccagc taagtttagg tctaggcatg gtagtaaaaa cctcccatcc
     1741 caagagtcaa tgacaagcag atctctgagt tcaaggcaag cccagacaga gcaagttcta
     1801 ggtgaagaaa aacgtaaacc caggcttggt agaccatacc tttaatccca gtgcttagca
     1861 gattggcttg catttgtctg agttcaaagt cagtcttcag agcaagttcc aggacagcca
     1921 agcttaggca gtggaggagt tggaaaacag aaaggctgat gatagtgtag tagaacaagg
     1981 ggaccatgtt ccagtcccag caagcagcaa aacttggcag ctttagacac atggctctgg
     2041 ttttagagtc tagaatagaa gtgactactg ggccaattag tgctggttag ctagagataa
     2101 gaaattagtg gtgattaaga agagaccagc atcagtgagg taaaatattc tggaaaatat
     2161 tttctgagag cacaatgaag ctgtgttcca gagatagtca agggagctcg ggctgggcat
     2221 aaaagtcagg gcagagccat ctattgctta catttgcttc tggcgtggcc accatggctc
     2281 ctaagaagaa gaggaaggtg atgagccagt tcgacatcct gtgcaagacc ccccccaagg
     2341 tgctggtgcg gcagttcgtg gagagattcg agaggcccag cggcgagaag atcgccagct
     2401 gtgccgccga gctgacctac ctgtgctgga tgatcaccca caacggcacc gccatcaaga
     2461 gggccacctt catgagctac aacaccatca tcagcaacag cctgagcttc gacatcgtga
     2521 acaagagcct gcagttcaag tacaagaccc agaaggccac catcctggag gccagcctga
     2581 agaagctgat ccccgcctgg gagttcacca tcatccctta caacggccag aagcaccaga
     2641 gcgacatcac cgacatcgtg tccagcctgc agctgcagtt cgagagcagc gaggaggccg
     2701 acaagggcaa cagccacagc aagaagatgc tgaaggccct gctgtccgag ggcgagagca
     2761 tctgggagat caccgagaag atcctgaaca gcttcgagta caccagcagg ttcaccaaga
     2821 ccaagaccct gtaccagttc ctgttcctgg ccacattcat caactgcggc aggttcagcg
     2881 acatcaagaa cgtggacccc aagagcttca agctggtgca gaacaagtac ctgggcgtga
     2941 tcattcagtg cctggtgacc gagaccaaga caagcgtgtc caggcacatc tactttttca
     3001 gcgccagagg caggatcgac cccctggtgt acctggacga gttcctgagg aacagcgagc
     3061 ccgtgctgaa gagagtgaac aggaccggca acagcagcag caacaagcag gagtaccagc
     3121 tgctgaagga caacctggtg cgcagctaca acaaggccct gaagaagaac gccccctacc
     3181 ccatcttcgc tatcaagaac ggccctaaga gccacatcgg caggcacctg atgaccagct
     3241 ttctgagcat gaagggcctg accgagctga caaacgtggt gggcaactgg agcgacaaga
     3301 gggcctccgc cgtggccagg accacctaca cccaccagat caccgccatc cccgaccact
     3361 acttcgccct ggtgtccagg tactacgcct acgaccccat cagcaaggag atgatcgccc
     3421 tgaaggacga gaccaacccc atcgaggagt ggcagcacat cgagcagctg aagggcagcg
     3481 ccgagggcag catcagatac cccgcctgga acggcatcat cagccaggag gtgctggact
     3541 acctgagcag ctacatcaac aggcggatct gagaattcga tatcaagctt atcgataatc
     3601 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3661 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3721 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3781 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3841 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3901 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3961 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     4021 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4081 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4141 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     4201 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     4261 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     4321 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     4381 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     4441 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     4501 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     4561 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     4621 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     4681 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     4741 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4801 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4861 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4921 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     4981 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     5041 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5101 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5161 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5221 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     5281 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     5341 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     5401 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5461 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5521 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5581 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5641 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5701 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5761 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5821 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5881 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5941 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6001 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6061 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6121 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6181 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6241 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6301 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6361 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6421 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6481 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6541 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6601 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6661 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6721 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6781 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6841 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6901 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6961 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7021 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7081 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7141 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7201 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7261 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7321 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7381 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7441 tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca
     7501 ctgaggcc
//
