LOCUS       sequence-466226-        6566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2102m 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 6566)
  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 6566)
  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: AiP1251 - pAAV-AiE2102m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(91..110)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   408..548
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   408..537
                     /label=AAV2 ITR
     rep_origin      623..1078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(710..729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     920..941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1095..1114)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1214..1236
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1274..1292)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1360..1464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1465..1533,1534..2325)
                     /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(1683..1702)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2496..3084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2985..3004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3146..3275
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      4509..4518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4515..4520,4521..4541,4542..5813)
                     /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    5838..6426
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5891..5911)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6309..6320)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(6458..6566,1..368)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
ORIGIN
        1 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
       61 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
      121 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
      181 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
      241 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
      301 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
      361 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
      421 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
      481 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
      541 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      601 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      661 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
      721 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      781 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
      841 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      901 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
      961 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     1021 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     1081 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     1141 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     1201 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     1261 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     1321 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1381 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1441 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1501 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1561 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1621 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1681 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1741 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1801 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1861 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1921 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1981 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2041 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2101 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2161 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2221 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2281 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2341 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2401 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2461 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2521 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2581 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2641 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2701 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2761 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2821 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2881 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2941 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3001 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3061 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3121 ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag
     3181 gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     3241 agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgtt ggtcaaagct
     3301 tgtcaggaca acgttattta aagtgcccag tacgtggcac tttgggtgaa aaaatcagga
     3361 actttccatg agtccccctc tgctgattag ttttttaatc aacttgatac aaactaaaga
     3421 tacctcggaa gacagagcct cggttgagga tttcctttca tccaactacc ctgtagccaa
     3481 gcctgtggga catcttgact gatgatggat atgggaaggc ctatgtcact gtggctgatg
     3541 tcatccccaa gcattctggg ttatatacaa aatttagcca agcaagttgg gaagcaactc
     3601 tcccccatgg tctctactct agttcctgcc tttcttgaat tcctgtcctg acttccccag
     3661 gtgatgaact gtaacccata agatgaaata aagctttcct ccctgttact ataggtcatg
     3721 gcgtttacta cagcaataga aagcaaacta ggacagatac tggtgccaca gagaagggat
     3781 actgctgtga cagacctgac cgtgctttgg ggacagactg tggaagaatt tggaactttg
     3841 ggataggaaa gcgattggat actcagagct taatgttttt ctctgtggga acttggaaga
     3901 gaatgtttaa aaaatcagct accaaataca gacactattg tatatgccag caagattttg
     3961 ctgaaaggac cctgatatag ctgtctcttg tgaggctttg ccagtgcctg gcaaatacag
     4021 aagtggatgc tcacagtcat ctatcagatg gaacacaggg cccccaatgg agaagctaga
     4081 gaaagtatcc aaggagctga aggtgtctgc agccttatag gtggaacaac aatatgaact
     4141 aaccagtacc cctggagctc gggtctctag ctgcatgtgt agcagaagat ggcctagtcg
     4201 gccatcactg ggaagagagg tcccttggtc ttgcaaactt tatacgcccc agtacagggg
     4261 aatgccaggg ccaagaagcg ggagtgggtg ggttggggag cagggtaggg ggagagtata
     4321 ggggaccttt gggatagcat ttgaaatgta aatgaagaaa atatctaata aaaaatgcag
     4381 atagtggagg cctggctagt ccctcaaagg ctagatctgg gctgttcatg tgatatttct
     4441 taaggagctc gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt
     4501 ctggcgtggc caccatggct cctaagaaga agaggaaggt gatgagccag ttcgacatcc
     4561 tgtgcaagac cccccccaag gtgctggtgc ggcagttcgt ggagagattc gagaggccca
     4621 gcggcgagaa gatcgccagc tgtgccgccg agctgaccta cctgtgctgg atgatcaccc
     4681 acaacggcac cgccatcaag agggccacct tcatgagcta caacaccatc atcagcaaca
     4741 gcctgagctt cgacatcgtg aacaagagcc tgcagttcaa gtacaagacc cagaaggcca
     4801 ccatcctgga ggccagcctg aagaagctga tccccgcctg ggagttcacc atcatccctt
     4861 acaacggcca gaagcaccag agcgacatca ccgacatcgt gtccagcctg cagctgcagt
     4921 tcgagagcag cgaggaggcc gacaagggca acagccacag caagaagatg ctgaaggccc
     4981 tgctgtccga gggcgagagc atctgggaga tcaccgagaa gatcctgaac agcttcgagt
     5041 acaccagcag gttcaccaag accaagaccc tgtaccagtt cctgttcctg gccacattca
     5101 tcaactgcgg caggttcagc gacatcaaga acgtggaccc caagagcttc aagctggtgc
     5161 agaacaagta cctgggcgtg atcattcagt gcctggtgac cgagaccaag acaagcgtgt
     5221 ccaggcacat ctactttttc agcgccagag gcaggatcga ccccctggtg tacctggacg
     5281 agttcctgag gaacagcgag cccgtgctga agagagtgaa caggaccggc aacagcagca
     5341 gcaacaagca ggagtaccag ctgctgaagg acaacctggt gcgcagctac aacaaggccc
     5401 tgaagaagaa cgccccctac cccatcttcg ctatcaagaa cggccctaag agccacatcg
     5461 gcaggcacct gatgaccagc tttctgagca tgaagggcct gaccgagctg acaaacgtgg
     5521 tgggcaactg gagcgacaag agggcctccg ccgtggccag gaccacctac acccaccaga
     5581 tcaccgccat ccccgaccac tacttcgccc tggtgtccag gtactacgcc tacgacccca
     5641 tcagcaagga gatgatcgcc ctgaaggacg agaccaaccc catcgaggag tggcagcaca
     5701 tcgagcagct gaagggcagc gccgagggca gcatcagata ccccgcctgg aacggcatca
     5761 tcagccagga ggtgctggac tacctgagca gctacatcaa caggcggatc tgagaattcg
     5821 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5881 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5941 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6001 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6061 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6121 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6181 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6241 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     6301 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6361 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6421 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     6481 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     6541 aaaattaagt tgcatcattt tgtctg
//
