LOCUS       sequence-466220-        5916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2067m 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 5916)
  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 5916)
  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: AiP1239 - pAAV-AiE2067m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     559..578
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(797..901)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     969..987
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1025..1047)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1147..1166
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1183..1638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1320..1341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1532..1551
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1713..1853
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1724..1853
                     /label=AAV2 ITR
     polyA_signal    1893..2369
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2151..2170
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2401..2989
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2507..2518
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2916..2936
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3014..4285,4286..4306,4307..4312))
                     /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"
     regulatory      4309..4318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4902..5031
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5093..5681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5173..5192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5852..5916,1..727,728..796))
                     /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"
ORIGIN
        1 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
       61 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      121 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      181 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
      241 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      301 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      361 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      421 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
      481 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
      541 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
      601 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
      661 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
      721 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
      781 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
      841 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
      901 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
      961 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     1021 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     1081 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     1141 actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata
     1201 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg
     1261 aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc
     1321 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1381 ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt
     1441 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1501 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1561 aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1621 cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag
     1681 atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct
     1741 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     1801 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     1861 ctcggtccgc acgtggttac ctacaaaatc agaaggacag ggaagggagc agtggttcac
     1921 gcctgtaatc ccagcaattt gggaggccaa ggtgggtaga tcacctgaga ttaggagttg
     1981 gagaccagcc tggccaatat ggtgaaaccc cgtctctacc aaaaaaacaa aaattagctg
     2041 agcctggtca tgcatgcctg gaatcccaac aactcgggag gctgaggcag gagaatcgct
     2101 tgaacccagg aggcggagat tgcagtgagc caagattgtg ccactgcact ccagcttggt
     2161 tcccaataga ccccgcaggc cctacaggtt gtcttcccaa cttgcccctt gctccatacc
     2221 acccccctcc accccataat attatagaag gacacctagt cagacaaaat gatgcaactt
     2281 aattttatta ggacaaggct ggtgggcact ggagtggcaa cttccagggc caggagaggc
     2341 actggggagg ggtcacaggg atgccacccg tagatctctc gagcagcgct cggtatcgat
     2401 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
     2461 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
     2521 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacataggcg agcagccaag
     2581 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
     2641 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
     2701 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
     2761 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
     2821 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     2881 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     2941 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tcgataagct
     3001 tgatatcgaa ttctcagatc cgcctgttga tgtagctgct caggtagtcc agcacctcct
     3061 ggctgatgat gccgttccag gcggggtatc tgatgctgcc ctcggcgctg cccttcagct
     3121 gctcgatgtg ctgccactcc tcgatggggt tggtctcgtc cttcagggcg atcatctcct
     3181 tgctgatggg gtcgtaggcg tagtacctgg acaccagggc gaagtagtgg tcggggatgg
     3241 cggtgatctg gtgggtgtag gtggtcctgg ccacggcgga ggccctcttg tcgctccagt
     3301 tgcccaccac gtttgtcagc tcggtcaggc ccttcatgct cagaaagctg gtcatcaggt
     3361 gcctgccgat gtggctctta gggccgttct tgatagcgaa gatggggtag ggggcgttct
     3421 tcttcagggc cttgttgtag ctgcgcacca ggttgtcctt cagcagctgg tactcctgct
     3481 tgttgctgct gctgttgccg gtcctgttca ctctcttcag cacgggctcg ctgttcctca
     3541 ggaactcgtc caggtacacc agggggtcga tcctgcctct ggcgctgaaa aagtagatgt
     3601 gcctggacac gcttgtcttg gtctcggtca ccaggcactg aatgatcacg cccaggtact
     3661 tgttctgcac cagcttgaag ctcttggggt ccacgttctt gatgtcgctg aacctgccgc
     3721 agttgatgaa tgtggccagg aacaggaact ggtacagggt cttggtcttg gtgaacctgc
     3781 tggtgtactc gaagctgttc aggatcttct cggtgatctc ccagatgctc tcgccctcgg
     3841 acagcagggc cttcagcatc ttcttgctgt ggctgttgcc cttgtcggcc tcctcgctgc
     3901 tctcgaactg cagctgcagg ctggacacga tgtcggtgat gtcgctctgg tgcttctggc
     3961 cgttgtaagg gatgatggtg aactcccagg cggggatcag cttcttcagg ctggcctcca
     4021 ggatggtggc cttctgggtc ttgtacttga actgcaggct cttgttcacg atgtcgaagc
     4081 tcaggctgtt gctgatgatg gtgttgtagc tcatgaaggt ggccctcttg atggcggtgc
     4141 cgttgtgggt gatcatccag cacaggtagg tcagctcggc ggcacagctg gcgatcttct
     4201 cgccgctggg cctctcgaat ctctccacga actgccgcac cagcaccttg gggggggtct
     4261 tgcacaggat gtcgaactgg ctcatcacct tcctcttctt cttaggagcc atggtggcca
     4321 cgccagaagc aaatgtaagc aatagatggc tctgccctga cttttatgcc cagcccgagc
     4381 tcctgtattg acaattacct ccagtgttgc atggaattta atgcctaaat aatgaagtca
     4441 aagaacaatg agggagactg agcatttgaa tatacttgat tgattcattt ctcgatcccg
     4501 tttaataacc cacttatttg gtgacactta agttcagcag acttagaaag aacagaacac
     4561 gccagagaga caatgttgca cagattaata actggatgtg ggtacaggct agtgccccct
     4621 aatagcagca caaaaatagg ttatcgtcaa aaatctagac ttaatgttct gaaatgggga
     4681 ttcgtgcttt tcctttatat gagtccatac ttatcaccat gggctaatgg gacataaacc
     4741 agaaatcagg agctaactgt gcaattccgg gggagtgggc tttttgaatt taataccttc
     4801 taagatcgtg atctttgctt gagatcttta atgtttaaat tacaatgtgt gaatattaaa
     4861 cacagttgat tgcaactgaa ctctctgacg cgtgcggccg caggaacccc tagtgatgga
     4921 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4981 ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag
     5041 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5101 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5161 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5221 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5281 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5341 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5401 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5461 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5521 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5581 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5641 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5701 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5761 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5821 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5881 tctcagcgat ctgtctattt cgttcatcca tagttg
//
