LOCUS       sequence-461951-        6348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2100m 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 6348)
  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 6348)
  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: AiP1249 - pAAV-AiE2100m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1146..1155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1152..1157,1158..1178,1179..2450)
                     /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    2475..3063
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2528..2548)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2946..2957)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3095..3571
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3294..3313)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3611..3751
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3611..3740
                     /label=AAV2 ITR
     rep_origin      3826..4281
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3913..3932)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4123..4144
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4298..4317)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4417..4439
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4477..4495)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4563..4667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4668..4736,4737..5528)
                     /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(4886..4905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5699..6287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6188..6207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcgtcaa acacagctca agctgacctt gatttgcttg
      181 taatcctcct attttggctt tctggatgct aggacaaagg catgagccac cagccctagt
      241 aagattaggc tttgttttgt ttgtatttcc ttgacttagc aattagaaag cctaaaagaa
      301 accacttgtg atcaaattca atgtgcttcc aaagttcgct gaggatcaga tgtcgtgggg
      361 atggcagact tcaaacctgt ttctgaacta ttttgctgtg tctttatgtc ccaggtcaaa
      421 ggatttcaac tgatgctcca taaagcaaga agggaaaatg gatttcttaa tgagtctgta
      481 tacagtgcag tatctatgag ctatggtgtg tgtgactgcc aaggcagtca gatgctattc
      541 aagcaaggaa tgctgctctg agatcagcta gtacgtaaga aagacatcct ggctcctgga
      601 agatgacaag agtttgcctc cgtagtattt gaaagaaagt ggtcatggcc atcatgaatc
      661 aatcacttca ggaaaaggtt gagcatgtgc tgtgacgact gcagatcttt ttatagctgt
      721 ggattctttt atagctgctg agatacgacg ataggagtgg agcggtagtt atcgttgatt
      781 tgtttggtaa tgtttcaggc aggtctacga attcctttgg attgtgggtt gtctcccaat
      841 cctacacttt cctagctctc taagaattcc cagagggtat ggagccagta gagactgacc
      901 acaaaagttg acacttaatg ttggctgacc tgcaaagaag gtttcattat gcattctttt
      961 ctgcacatgt gaaatcagag accatgggat ccatttcaat aacaaagccc atctttgtaa
     1021 agtagaagct agttgaaaat taagtatcat tttctattgt ggtgattggg aagaaatatt
     1081 ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     1141 gcgtggccac catggctcct aagaagaaga ggaaggtgat gagccagttc gacatcctgt
     1201 gcaagacccc ccccaaggtg ctggtgcggc agttcgtgga gagattcgag aggcccagcg
     1261 gcgagaagat cgccagctgt gccgccgagc tgacctacct gtgctggatg atcacccaca
     1321 acggcaccgc catcaagagg gccaccttca tgagctacaa caccatcatc agcaacagcc
     1381 tgagcttcga catcgtgaac aagagcctgc agttcaagta caagacccag aaggccacca
     1441 tcctggaggc cagcctgaag aagctgatcc ccgcctggga gttcaccatc atcccttaca
     1501 acggccagaa gcaccagagc gacatcaccg acatcgtgtc cagcctgcag ctgcagttcg
     1561 agagcagcga ggaggccgac aagggcaaca gccacagcaa gaagatgctg aaggccctgc
     1621 tgtccgaggg cgagagcatc tgggagatca ccgagaagat cctgaacagc ttcgagtaca
     1681 ccagcaggtt caccaagacc aagaccctgt accagttcct gttcctggcc acattcatca
     1741 actgcggcag gttcagcgac atcaagaacg tggaccccaa gagcttcaag ctggtgcaga
     1801 acaagtacct gggcgtgatc attcagtgcc tggtgaccga gaccaagaca agcgtgtcca
     1861 ggcacatcta ctttttcagc gccagaggca ggatcgaccc cctggtgtac ctggacgagt
     1921 tcctgaggaa cagcgagccc gtgctgaaga gagtgaacag gaccggcaac agcagcagca
     1981 acaagcagga gtaccagctg ctgaaggaca acctggtgcg cagctacaac aaggccctga
     2041 agaagaacgc cccctacccc atcttcgcta tcaagaacgg ccctaagagc cacatcggca
     2101 ggcacctgat gaccagcttt ctgagcatga agggcctgac cgagctgaca aacgtggtgg
     2161 gcaactggag cgacaagagg gcctccgccg tggccaggac cacctacacc caccagatca
     2221 ccgccatccc cgaccactac ttcgccctgg tgtccaggta ctacgcctac gaccccatca
     2281 gcaaggagat gatcgccctg aaggacgaga ccaaccccat cgaggagtgg cagcacatcg
     2341 agcagctgaa gggcagcgcc gagggcagca tcagataccc cgcctggaac ggcatcatca
     2401 gccaggaggt gctggactac ctgagcagct acatcaacag gcggatctga gaattcgata
     2461 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2521 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2581 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2641 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2701 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2761 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2821 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     2881 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2941 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3001 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3061 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     3121 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     3181 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     3241 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     3301 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     3361 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     3421 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     3481 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     3541 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     3601 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3661 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3721 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3781 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3841 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3901 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3961 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4021 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4081 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4141 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     4201 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4261 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4321 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4381 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4441 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4501 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4561 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4681 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4741 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4801 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4861 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4921 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4981 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5041 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5101 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5161 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5221 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5281 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5341 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5401 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5461 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5521 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5581 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5641 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5701 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5761 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5821 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5881 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5941 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6001 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6061 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6121 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6181 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6241 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6301 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
