LOCUS       sequence-466227-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2103m 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 6992)
  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 6992)
  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: AiP1252 - pAAV-AiE2103m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      797..806
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(803..808,809..829,830..2101)
                     /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    2126..2714
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2179..2199)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2597..2608)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2746..3222
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2945..2964)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3262..3402
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3262..3391
                     /label=AAV2 ITR
     rep_origin      3477..3932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3564..3583)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3774..3795
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3949..3968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4068..4090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4128..4146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4214..4318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4319..4387,4388..5179)
                     /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(4537..4556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5350..5938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5839..5858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6000..6129
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ttttactctt ctctccttgt ggctactccc ttccactgcc accgccactt cctgtcttat
       61 ccaggcaatc atccagattt gtgcaaacaa gagttttcac aagtgaactc ttccaactag
      121 atctttaggc aaattacttc agcaataact tttaaatcaa attaatatct caaacctaaa
      181 cattctacag gccatggggt taacctgaaa actcaacagt accttttaat gttctggcta
      241 gatctgaaac tcacgtttta gtaaaataaa atcattagtt ggtataccct acataataga
      301 attgtttttt tattctacat taacttcaga gcctcttctg aaatataatg atgtctttca
      361 tgctttattg tgattaatac aaaaactgga ttaaacatat ccatatacaa cttcttggct
      421 tcatattagg cctgaatgca gaactggcag cacatagcag aacaattacc aacagaaaag
      481 tccccttttt caaagttcat tttgtgtttc cctttcatta cacaaaagat tagataacaa
      541 agagttttct ttgctacaaa aaattatctt tcttcagcat tataactgat ggagaaaatt
      601 actttggtga aaatgaaaat agaaagcata agtgttaata gatgaattca tatctaatta
      661 tctacttctc gcttgcaaaa aaagaaatag tgggatactt aagttaaaat gttctgccca
      721 gaaatcaaag acgagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
      781 atttgcttct ggcgtggcca ccatggctcc taagaagaag aggaaggtga tgagccagtt
      841 cgacatcctg tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg agagattcga
      901 gaggcccagc ggcgagaaga tcgccagctg tgccgccgag ctgacctacc tgtgctggat
      961 gatcacccac aacggcaccg ccatcaagag ggccaccttc atgagctaca acaccatcat
     1021 cagcaacagc ctgagcttcg acatcgtgaa caagagcctg cagttcaagt acaagaccca
     1081 gaaggccacc atcctggagg ccagcctgaa gaagctgatc cccgcctggg agttcaccat
     1141 catcccttac aacggccaga agcaccagag cgacatcacc gacatcgtgt ccagcctgca
     1201 gctgcagttc gagagcagcg aggaggccga caagggcaac agccacagca agaagatgct
     1261 gaaggccctg ctgtccgagg gcgagagcat ctgggagatc accgagaaga tcctgaacag
     1321 cttcgagtac accagcaggt tcaccaagac caagaccctg taccagttcc tgttcctggc
     1381 cacattcatc aactgcggca ggttcagcga catcaagaac gtggacccca agagcttcaa
     1441 gctggtgcag aacaagtacc tgggcgtgat cattcagtgc ctggtgaccg agaccaagac
     1501 aagcgtgtcc aggcacatct actttttcag cgccagaggc aggatcgacc ccctggtgta
     1561 cctggacgag ttcctgagga acagcgagcc cgtgctgaag agagtgaaca ggaccggcaa
     1621 cagcagcagc aacaagcagg agtaccagct gctgaaggac aacctggtgc gcagctacaa
     1681 caaggccctg aagaagaacg ccccctaccc catcttcgct atcaagaacg gccctaagag
     1741 ccacatcggc aggcacctga tgaccagctt tctgagcatg aagggcctga ccgagctgac
     1801 aaacgtggtg ggcaactgga gcgacaagag ggcctccgcc gtggccagga ccacctacac
     1861 ccaccagatc accgccatcc ccgaccacta cttcgccctg gtgtccaggt actacgccta
     1921 cgaccccatc agcaaggaga tgatcgccct gaaggacgag accaacccca tcgaggagtg
     1981 gcagcacatc gagcagctga agggcagcgc cgagggcagc atcagatacc ccgcctggaa
     2041 cggcatcatc agccaggagg tgctggacta cctgagcagc tacatcaaca ggcggatctg
     2101 agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     2161 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2221 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2281 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2341 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2401 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2461 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2521 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     2581 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2641 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2701 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt
     2761 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     2821 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     2881 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     2941 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     3001 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     3061 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     3121 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     3181 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     3241 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     3301 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3361 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     3421 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     3481 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3541 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3601 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3661 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     3721 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3781 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     3841 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3901 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3961 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     4021 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4081 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     4141 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4201 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4261 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4321 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4381 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4441 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4501 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4561 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4621 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4681 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4741 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4801 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4861 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4921 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4981 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5041 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5101 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5161 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5221 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5281 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5341 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5401 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5461 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5521 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5581 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5641 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5701 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5761 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5821 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5881 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5941 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc
     6001 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
     6061 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     6121 ggggttcctg cggccgcacg cgtgcaaagg atatctgcag tgtcactcaa agcttcagtc
     6181 tgaataggaa atgtctcaac aggttgtagt tagtgataac ttagtctcca gttagtgatg
     6241 ctgttttaga agacttttag acctttagaa gaaggagcct tgcttaatag agtgggtcac
     6301 aaatggaaaa ttttgatgtt ttgtgctctg acccccattt ttcagttcac tctccatttc
     6361 ctgtctttgg acacagtatg accagccagc ctccttctcc tgctgaagtg cattctgttt
     6421 tatattttca agagaatgat ctatgctagc ccaacgttca gttttgaatc aattgagtta
     6481 atttggtata aaaagtaaga caaagtaaag tagaataaca accagcagac agaaactcag
     6541 gaaaagaatg agaagatgaa tatagaacac tattatacac atgtattccc tagcaatcac
     6601 tagacaatgt ccaaagctct acaatagtgt ggcaagactc tacaagagcc agaaaacaca
     6661 attctaaaga gctgaacaca aacatcagca ccatgagcga tggaggaacc aagtgtttct
     6721 ttgagatgct agtagagtga agacatacca gcaaacatca caaaaatgta aaataagttg
     6781 caggggtggg caggacttta gggattttga ttgttggtta ttgctttttc tgaaatactt
     6841 aaaagaatga cttaaaattc ctgttcattg cacattgttt ctgggctgag tgtgtacatt
     6901 tctacatgtg atatagggtc tcagaaacac ttccaggaca ctgtacacct ttcagtttgg
     6961 tatttatcca tggaatttta attgagaaaa ac
//
