LOCUS       sequence-466202-        6021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2037m 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 6021)
  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 6021)
  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: AiP1174 - pAAV-AiE2037m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(213..1004,1005..1073))
                     /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     836..855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1074..1178)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1246..1264
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1302..1324)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1424..1443
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1460..1915)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1597..1618)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1809..1828
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1990..2130
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2001..2130
                     /label=AAV2 ITR
     polyA_signal    2170..2646
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2428..2447
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2678..3266
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2784..2795
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3193..3213
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3291..4562,4563..4583,4584..4589))
                     /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      4586..4595
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   5284..5413
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(5475..6021,1..42))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5555..5574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
       61 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
      361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
      721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1201 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1261 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1321 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1381 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     1441 gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     1501 ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa
     1561 tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta
     1621 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1681 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1741 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1801 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc
     1861 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat
     1921 ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     1981 tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
     2041 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2101 gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg tggttaccta
     2161 caaaatcaga aggacaggga agggagcagt ggttcacgcc tgtaatccca gcaatttggg
     2221 aggccaaggt gggtagatca cctgagatta ggagttggag accagcctgg ccaatatggt
     2281 gaaaccccgt ctctaccaaa aaaacaaaaa ttagctgagc ctggtcatgc atgcctggaa
     2341 tcccaacaac tcgggaggct gaggcaggag aatcgcttga acccaggagg cggagattgc
     2401 agtgagccaa gattgtgcca ctgcactcca gcttggttcc caatagaccc cgcaggccct
     2461 acaggttgtc ttcccaactt gccccttgct ccataccacc cccctccacc ccataatatt
     2521 atagaaggac acctagtcag acaaaatgat gcaacttaat tttattagga caaggctggt
     2581 gggcactgga gtggcaactt ccagggccag gagaggcact ggggaggggt cacagggatg
     2641 ccacccgtag atctctcgag cagcgctcgg tatcgatgcg gggaggcggc ccaaagggag
     2701 atccgactcg tctgagggcg aaggcgaaga cgcggaagag gccgcagagc cggcagcagg
     2761 ccgcgggaag gaaggtccgc tggattgagg gccgaaggga cgtagcagaa ggacgtcccg
     2821 cgcagaatcc aggtggcaac ataggcgagc agccaaggaa aggacgatga tttccccgac
     2881 aacaccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag
     2941 gcggcgatga gttccgccgt ggcaataggg agggggaaag cgaaagtccc ggaaaggagc
     3001 tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc
     3061 acgccacgtt gcctgacaac gggccacaac tcctcataaa gagacagcaa ccaggattta
     3121 tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa
     3181 gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca
     3241 caaattttgt aatccagagg ttgattatcg ataagcttga tatcgaattc tcagatccgc
     3301 ctgttgatgt agctgctcag gtagtccagc acctcctggc tgatgatgcc gttccaggcg
     3361 gggtatctga tgctgccctc ggcgctgccc ttcagctgct cgatgtgctg ccactcctcg
     3421 atggggttgg tctcgtcctt cagggcgatc atctccttgc tgatggggtc gtaggcgtag
     3481 tacctggaca ccagggcgaa gtagtggtcg gggatggcgg tgatctggtg ggtgtaggtg
     3541 gtcctggcca cggcggaggc cctcttgtcg ctccagttgc ccaccacgtt tgtcagctcg
     3601 gtcaggccct tcatgctcag aaagctggtc atcaggtgcc tgccgatgtg gctcttaggg
     3661 ccgttcttga tagcgaagat ggggtagggg gcgttcttct tcagggcctt gttgtagctg
     3721 cgcaccaggt tgtccttcag cagctggtac tcctgcttgt tgctgctgct gttgccggtc
     3781 ctgttcactc tcttcagcac gggctcgctg ttcctcagga actcgtccag gtacaccagg
     3841 gggtcgatcc tgcctctggc gctgaaaaag tagatgtgcc tggacacgct tgtcttggtc
     3901 tcggtcacca ggcactgaat gatcacgccc aggtacttgt tctgcaccag cttgaagctc
     3961 ttggggtcca cgttcttgat gtcgctgaac ctgccgcagt tgatgaatgt ggccaggaac
     4021 aggaactggt acagggtctt ggtcttggtg aacctgctgg tgtactcgaa gctgttcagg
     4081 atcttctcgg tgatctccca gatgctctcg ccctcggaca gcagggcctt cagcatcttc
     4141 ttgctgtggc tgttgccctt gtcggcctcc tcgctgctct cgaactgcag ctgcaggctg
     4201 gacacgatgt cggtgatgtc gctctggtgc ttctggccgt tgtaagggat gatggtgaac
     4261 tcccaggcgg ggatcagctt cttcaggctg gcctccagga tggtggcctt ctgggtcttg
     4321 tacttgaact gcaggctctt gttcacgatg tcgaagctca ggctgttgct gatgatggtg
     4381 ttgtagctca tgaaggtggc cctcttgatg gcggtgccgt tgtgggtgat catccagcac
     4441 aggtaggtca gctcggcggc acagctggcg atcttctcgc cgctgggcct ctcgaatctc
     4501 tccacgaact gccgcaccag caccttgggg ggggtcttgc acaggatgtc gaactggctc
     4561 atcaccttcc tcttcttctt aggagccatg gtggccacgc cagaagcaaa tgtaagcaat
     4621 agatggctct gccctgactt ttatgcccag cccgagctcc ttaaggacat tggcatacat
     4681 ctatcacaac cacaatcctt ttacttcttc tcaactctgt ccttctccac agtaactaat
     4741 tacacttcca gctatttcag taatagcttt ttgttttctt gatcatctca tcaccaaaat
     4801 gtttgtgatt aagcagcaga ttcattatcc ttcattcatc tctaaggctc tttttatcta
     4861 gagcatttta ttttcactta taatggatgt gttcaagaag tactattgct tgttctctaa
     4921 gaccttccca gaattgattt cgcttgatgg catctcctat tgtcatttga tgtgctactt
     4981 tgtctacact tgctgtaaat tggtacttag ttctgtaaga ttagatttgt gttgtttttt
     5041 ttttccttta tttttgcttc atagattgtg aagtgttcta tttggagatg tataaagttt
     5101 gagtgtctct ttttatctat gacattaggt gccattaatg atcaataccc ttgttttcta
     5161 tgctaatcat gcctttatag atatttttgt tctcagatgt tcttcctcta aaataaagaa
     5221 taagttgttg aaataatatc tctcatgctt tcgtactctc atgattgtca cgcgtgcggc
     5281 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     5341 aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc
     5401 agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5461 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5521 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5581 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5641 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5701 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5761 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5821 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5881 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5941 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6001 gctggtagcg gtggtttttt t
//
