LOCUS       sequence-483861-        6109 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  AiE0082h 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 6109)
  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 6109)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 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: AiP1231 - pAAV-AiE0082h-minBG-FlpO-WPRE-HGHpA
            ...
FEATURES             Location/Qualifiers
     source          1..6109
                     /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      907..916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(913..918,919..939,940..2211)
                     /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    2236..2824
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2289..2309)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2707..2718)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2856..3332
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3055..3074)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3372..3512
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3372..3501
                     /label=AAV2 ITR
     rep_origin      3587..4042
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3674..3693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3884..3905
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4059..4078)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4178..4200
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4238..4256)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4324..4428
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4429..4497,4498..5289)
                     /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(4647..4666)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5460..6048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5949..5968
                     /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 gcgtcctttg gccggcagga gcagcctgat tttagtgact
      181 cgttcagaag ggaggtgcag ctggaaggtt gtgtttatta aagcccaagt gactggaggt
      241 aaatgcaggt gtgttcagga aatgtagctg gttcacctct cagtaggcac aaggccaaag
      301 tcagccgact tgcagattgt aatgccggtt ctcccaccca gaggttattt gagttcatcc
      361 agcactgcag ctgagcagaa tgctgcagtc agggtgacac aatgcaaact ggagccatgt
      421 cctcaagggc atgccaatca aaccaacttt aaccttcgct ggtttgtggc tgctgaccta
      481 gagaggaatt tgttatatgg ctgaggtcct gtcctaccta agccagactc cagtgcaaaa
      541 aaaaatcctg tcagatgcgc ctcttcttct ctgtctccat ctccctctct gcctcccgtt
      601 tcccctttac tctttactgt tagttggcct ggggttcctg ggctgaagtg gagtttcctc
      661 ctgtctccgt gtgtttgctg ctctggtttg ctcttcttcc agggcccctt ggtaaaagaa
      721 gagctgctga cagggtagag tggaggtggg gaagtgacca ccagactggg agtgatgtag
      781 gttgtggtca tcaatgatgt aggttgctgt caccaggaca ctcaaatcag ctgtgaccag
      841 aggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
      901 ggcgtggcca ccatggctcc taagaagaag aggaaggtga tgagccagtt cgacatcctg
      961 tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg agagattcga gaggcccagc
     1021 ggcgagaaga tcgccagctg tgccgccgag ctgacctacc tgtgctggat gatcacccac
     1081 aacggcaccg ccatcaagag ggccaccttc atgagctaca acaccatcat cagcaacagc
     1141 ctgagcttcg acatcgtgaa caagagcctg cagttcaagt acaagaccca gaaggccacc
     1201 atcctggagg ccagcctgaa gaagctgatc cccgcctggg agttcaccat catcccttac
     1261 aacggccaga agcaccagag cgacatcacc gacatcgtgt ccagcctgca gctgcagttc
     1321 gagagcagcg aggaggccga caagggcaac agccacagca agaagatgct gaaggccctg
     1381 ctgtccgagg gcgagagcat ctgggagatc accgagaaga tcctgaacag cttcgagtac
     1441 accagcaggt tcaccaagac caagaccctg taccagttcc tgttcctggc cacattcatc
     1501 aactgcggca ggttcagcga catcaagaac gtggacccca agagcttcaa gctggtgcag
     1561 aacaagtacc tgggcgtgat cattcagtgc ctggtgaccg agaccaagac aagcgtgtcc
     1621 aggcacatct actttttcag cgccagaggc aggatcgacc ccctggtgta cctggacgag
     1681 ttcctgagga acagcgagcc cgtgctgaag agagtgaaca ggaccggcaa cagcagcagc
     1741 aacaagcagg agtaccagct gctgaaggac aacctggtgc gcagctacaa caaggccctg
     1801 aagaagaacg ccccctaccc catcttcgct atcaagaacg gccctaagag ccacatcggc
     1861 aggcacctga tgaccagctt tctgagcatg aagggcctga ccgagctgac aaacgtggtg
     1921 ggcaactgga gcgacaagag ggcctccgcc gtggccagga ccacctacac ccaccagatc
     1981 accgccatcc ccgaccacta cttcgccctg gtgtccaggt actacgccta cgaccccatc
     2041 agcaaggaga tgatcgccct gaaggacgag accaacccca tcgaggagtg gcagcacatc
     2101 gagcagctga agggcagcgc cgagggcagc atcagatacc ccgcctggaa cggcatcatc
     2161 agccaggagg tgctggacta cctgagcagc tacatcaaca ggcggatctg agaattcgat
     2221 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2281 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2341 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2401 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2461 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2521 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2581 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2641 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2701 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2761 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2821 ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     2881 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     2941 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     3001 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     3061 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     3121 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     3181 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     3241 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     3301 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac
     3361 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3421 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3481 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     3541 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     3601 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3661 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3721 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3781 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     3841 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3901 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     3961 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4021 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     4081 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4141 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4201 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4261 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4321 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4381 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4441 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4501 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4561 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4621 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4681 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4741 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4801 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4861 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4921 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4981 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5041 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5101 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5161 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5221 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5281 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5341 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5401 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5461 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5521 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5581 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5641 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5701 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5761 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5821 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5881 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5941 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6001 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6061 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
