LOCUS       sequence-461955-        5921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2034m 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 5921)
  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 5921)
  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: AiP1171 - pAAV-AiE2034m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..5921
                     /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      719..728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(725..730,731..751,752..2023)
                     /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    2048..2636
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2101..2121)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2519..2530)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2668..3144
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2867..2886)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3184..3324
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3184..3313
                     /label=AAV2 ITR
     rep_origin      3399..3854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3486..3505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3696..3717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3871..3890)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3990..4012
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4050..4068)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4136..4240
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4241..4309,4310..5101)
                     /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(4459..4478)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5272..5860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5761..5780
                     /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 gcgtggagag aactatacca ctaatcctta ctaccccccc
      181 cccaattaat gacaccccat tcgagcatct ccaatgaaag gcgcattatt aatattcaag
      241 gaaaaaaacc cttgtaacca ttttagcaga tacagctaac aatgaactaa atttgctttg
      301 tgcaagacag agagtgagca tcccaaatta gtttatgtaa aagctgaaat cctcccctga
      361 aaggtctcag taccatcctc aaagaatatc ccgatcatca gcagtgccgt caagattccc
      421 gccaacatta tcccctttgt tgtctgaaat gtagaatcaa gcacatctgg caaatgtcct
      481 ggagctcaca aatctctcaa acggcgtcta attcccagtg gtgaaagaga gcgccactgc
      541 ctaatcgtgc ttctcttcaa aacagcactg actttccttc aatcagaaac aatggctttg
      601 ctttcttaaa aatatcacac gccagtgatt atttgcagag aggcccatct taaggagctc
      661 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctggcgtggc
      721 caccatggct cctaagaaga agaggaaggt gatgagccag ttcgacatcc tgtgcaagac
      781 cccccccaag gtgctggtgc ggcagttcgt ggagagattc gagaggccca gcggcgagaa
      841 gatcgccagc tgtgccgccg agctgaccta cctgtgctgg atgatcaccc acaacggcac
      901 cgccatcaag agggccacct tcatgagcta caacaccatc atcagcaaca gcctgagctt
      961 cgacatcgtg aacaagagcc tgcagttcaa gtacaagacc cagaaggcca ccatcctgga
     1021 ggccagcctg aagaagctga tccccgcctg ggagttcacc atcatccctt acaacggcca
     1081 gaagcaccag agcgacatca ccgacatcgt gtccagcctg cagctgcagt tcgagagcag
     1141 cgaggaggcc gacaagggca acagccacag caagaagatg ctgaaggccc tgctgtccga
     1201 gggcgagagc atctgggaga tcaccgagaa gatcctgaac agcttcgagt acaccagcag
     1261 gttcaccaag accaagaccc tgtaccagtt cctgttcctg gccacattca tcaactgcgg
     1321 caggttcagc gacatcaaga acgtggaccc caagagcttc aagctggtgc agaacaagta
     1381 cctgggcgtg atcattcagt gcctggtgac cgagaccaag acaagcgtgt ccaggcacat
     1441 ctactttttc agcgccagag gcaggatcga ccccctggtg tacctggacg agttcctgag
     1501 gaacagcgag cccgtgctga agagagtgaa caggaccggc aacagcagca gcaacaagca
     1561 ggagtaccag ctgctgaagg acaacctggt gcgcagctac aacaaggccc tgaagaagaa
     1621 cgccccctac cccatcttcg ctatcaagaa cggccctaag agccacatcg gcaggcacct
     1681 gatgaccagc tttctgagca tgaagggcct gaccgagctg acaaacgtgg tgggcaactg
     1741 gagcgacaag agggcctccg ccgtggccag gaccacctac acccaccaga tcaccgccat
     1801 ccccgaccac tacttcgccc tggtgtccag gtactacgcc tacgacccca tcagcaagga
     1861 gatgatcgcc ctgaaggacg agaccaaccc catcgaggag tggcagcaca tcgagcagct
     1921 gaagggcagc gccgagggca gcatcagata ccccgcctgg aacggcatca tcagccagga
     1981 ggtgctggac tacctgagca gctacatcaa caggcggatc tgagaattcg atatcaagct
     2041 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2101 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2161 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2221 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2281 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2341 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2401 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2461 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2521 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2581 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     2641 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     2701 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     2761 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     2821 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     2881 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     2941 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3001 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3061 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3121 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     3181 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3241 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3301 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     3361 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3421 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     3481 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3541 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3601 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3661 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3721 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3781 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3841 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3901 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3961 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4021 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4081 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4141 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4201 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4261 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4321 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4381 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4441 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4501 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4561 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4621 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4681 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4741 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4801 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4861 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4921 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4981 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5041 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5101 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5161 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5221 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5401 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5821 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5881 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
