LOCUS       sequence-466200-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0339m 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 5797)
  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 5797)
  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: AiP1100 - pAAV-AiE0339m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   18..158
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   18..147
                     /label=AAV2 ITR
     rep_origin      233..688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(320..339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     530..551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(705..724)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     824..846
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(884..902)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        970..1074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1075..1143,1144..1935)
                     /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(1293..1312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2106..2694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2595..2614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2756..2885
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3350..3359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3356..3361,3362..3382,3383..4654)
                     /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    4679..5267
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4732..4752)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5150..5161)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5299..5775
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5498..5517)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
       61 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
      121 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
      181 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
      241 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
      301 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
      361 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
      421 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
      481 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
      541 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
      601 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
      661 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
      721 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
      781 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
      841 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
      901 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
      961 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1021 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1081 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1141 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1201 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1261 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1321 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1381 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1441 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1501 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1561 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1621 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1681 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1741 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1801 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1861 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1921 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1981 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2041 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2101 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2161 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2221 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2281 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2341 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2401 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2461 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2521 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2581 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2641 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2701 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
     2761 aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg
     2821 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     2881 ttcctgcggc cgcacgcgta cacaaggaca agctggtgag cgctgttaaa tctctactac
     2941 aagctgacac tgtcacaccc acagatacac cggagtgtca gatggcccca tctcatggta
     3001 catgaaatgc agtaatcctg cggtgcaatt ttctgagcta ttttccagag ctttctcttg
     3061 aaaagcaggt gacttcttgg gtagcagcaa tttacctttc acaccgggta atcatcaagg
     3121 tgcctgattg ccctgaatac ctttcccggc tgtcagaata cttcaaggct gagaattagc
     3181 atttcaaaac cttttcaaat gagggtaggg gggctcattc ccttggttga aggttcccca
     3241 ggctgcaaag aagcaagtgt taagtgcaca cctggggaag ccatagagct cgggctgggc
     3301 ataaaagtca gggcagagcc atctattgct tacatttgct tctggcgtgg ccaccatggc
     3361 tcctaagaag aagaggaagg tgatgagcca gttcgacatc ctgtgcaaga ccccccccaa
     3421 ggtgctggtg cggcagttcg tggagagatt cgagaggccc agcggcgaga agatcgccag
     3481 ctgtgccgcc gagctgacct acctgtgctg gatgatcacc cacaacggca ccgccatcaa
     3541 gagggccacc ttcatgagct acaacaccat catcagcaac agcctgagct tcgacatcgt
     3601 gaacaagagc ctgcagttca agtacaagac ccagaaggcc accatcctgg aggccagcct
     3661 gaagaagctg atccccgcct gggagttcac catcatccct tacaacggcc agaagcacca
     3721 gagcgacatc accgacatcg tgtccagcct gcagctgcag ttcgagagca gcgaggaggc
     3781 cgacaagggc aacagccaca gcaagaagat gctgaaggcc ctgctgtccg agggcgagag
     3841 catctgggag atcaccgaga agatcctgaa cagcttcgag tacaccagca ggttcaccaa
     3901 gaccaagacc ctgtaccagt tcctgttcct ggccacattc atcaactgcg gcaggttcag
     3961 cgacatcaag aacgtggacc ccaagagctt caagctggtg cagaacaagt acctgggcgt
     4021 gatcattcag tgcctggtga ccgagaccaa gacaagcgtg tccaggcaca tctacttttt
     4081 cagcgccaga ggcaggatcg accccctggt gtacctggac gagttcctga ggaacagcga
     4141 gcccgtgctg aagagagtga acaggaccgg caacagcagc agcaacaagc aggagtacca
     4201 gctgctgaag gacaacctgg tgcgcagcta caacaaggcc ctgaagaaga acgcccccta
     4261 ccccatcttc gctatcaaga acggccctaa gagccacatc ggcaggcacc tgatgaccag
     4321 ctttctgagc atgaagggcc tgaccgagct gacaaacgtg gtgggcaact ggagcgacaa
     4381 gagggcctcc gccgtggcca ggaccaccta cacccaccag atcaccgcca tccccgacca
     4441 ctacttcgcc ctggtgtcca ggtactacgc ctacgacccc atcagcaagg agatgatcgc
     4501 cctgaaggac gagaccaacc ccatcgagga gtggcagcac atcgagcagc tgaagggcag
     4561 cgccgagggc agcatcagat accccgcctg gaacggcatc atcagccagg aggtgctgga
     4621 ctacctgagc agctacatca acaggcggat ctgagaattc gatatcaagc ttatcgataa
     4681 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4741 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4801 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4861 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4921 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4981 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5041 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     5101 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5161 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5221 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc
     5281 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     5341 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     5401 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     5461 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     5521 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     5581 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     5641 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     5701 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     5761 cccttccctg tccttctgat tttgtaggta accacgt
//
