LOCUS       sequence-466205-        6076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2041m 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 6076)
  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 6076)
  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: AiP1177 - pAAV-AiE2041m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(16..807,808..876))
                     /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     639..658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(877..981)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1049..1067
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1105..1127)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1227..1246
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1263..1718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1400..1421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1612..1631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1793..1933
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1804..1933
                     /label=AAV2 ITR
     polyA_signal    1973..2449
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2231..2250
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2481..3069
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2587..2598
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2996..3016
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3094..4365,4366..4386,4387..4392))
                     /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      4389..4398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   5142..5271
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5333..5921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5413..5432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
       61 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      121 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      181 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      241 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      301 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
      361 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      421 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      481 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      541 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
      601 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
      661 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
      721 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
      781 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
      841 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
      901 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
      961 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     1021 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     1081 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     1141 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     1201 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     1261 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1321 attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
     1381 gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     1441 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     1501 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     1561 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1621 agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1681 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
     1741 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg
     1801 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     1861 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     1921 actaggggtt cctgcggccg ctcggtccgc acgtggttac ctacaaaatc agaaggacag
     1981 ggaagggagc agtggttcac gcctgtaatc ccagcaattt gggaggccaa ggtgggtaga
     2041 tcacctgaga ttaggagttg gagaccagcc tggccaatat ggtgaaaccc cgtctctacc
     2101 aaaaaaacaa aaattagctg agcctggtca tgcatgcctg gaatcccaac aactcgggag
     2161 gctgaggcag gagaatcgct tgaacccagg aggcggagat tgcagtgagc caagattgtg
     2221 ccactgcact ccagcttggt tcccaataga ccccgcaggc cctacaggtt gtcttcccaa
     2281 cttgcccctt gctccatacc acccccctcc accccataat attatagaag gacacctagt
     2341 cagacaaaat gatgcaactt aattttatta ggacaaggct ggtgggcact ggagtggcaa
     2401 cttccagggc caggagaggc actggggagg ggtcacaggg atgccacccg tagatctctc
     2461 gagcagcgct cggtatcgat gcggggaggc ggcccaaagg gagatccgac tcgtctgagg
     2521 gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc
     2581 cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc
     2641 aacataggcg agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc
     2701 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
     2761 cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat
     2821 gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac
     2881 aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat
     2941 gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag
     3001 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
     3061 aggttgatta tcgataagct tgatatcgaa ttctcagatc cgcctgttga tgtagctgct
     3121 caggtagtcc agcacctcct ggctgatgat gccgttccag gcggggtatc tgatgctgcc
     3181 ctcggcgctg cccttcagct gctcgatgtg ctgccactcc tcgatggggt tggtctcgtc
     3241 cttcagggcg atcatctcct tgctgatggg gtcgtaggcg tagtacctgg acaccagggc
     3301 gaagtagtgg tcggggatgg cggtgatctg gtgggtgtag gtggtcctgg ccacggcgga
     3361 ggccctcttg tcgctccagt tgcccaccac gtttgtcagc tcggtcaggc ccttcatgct
     3421 cagaaagctg gtcatcaggt gcctgccgat gtggctctta gggccgttct tgatagcgaa
     3481 gatggggtag ggggcgttct tcttcagggc cttgttgtag ctgcgcacca ggttgtcctt
     3541 cagcagctgg tactcctgct tgttgctgct gctgttgccg gtcctgttca ctctcttcag
     3601 cacgggctcg ctgttcctca ggaactcgtc caggtacacc agggggtcga tcctgcctct
     3661 ggcgctgaaa aagtagatgt gcctggacac gcttgtcttg gtctcggtca ccaggcactg
     3721 aatgatcacg cccaggtact tgttctgcac cagcttgaag ctcttggggt ccacgttctt
     3781 gatgtcgctg aacctgccgc agttgatgaa tgtggccagg aacaggaact ggtacagggt
     3841 cttggtcttg gtgaacctgc tggtgtactc gaagctgttc aggatcttct cggtgatctc
     3901 ccagatgctc tcgccctcgg acagcagggc cttcagcatc ttcttgctgt ggctgttgcc
     3961 cttgtcggcc tcctcgctgc tctcgaactg cagctgcagg ctggacacga tgtcggtgat
     4021 gtcgctctgg tgcttctggc cgttgtaagg gatgatggtg aactcccagg cggggatcag
     4081 cttcttcagg ctggcctcca ggatggtggc cttctgggtc ttgtacttga actgcaggct
     4141 cttgttcacg atgtcgaagc tcaggctgtt gctgatgatg gtgttgtagc tcatgaaggt
     4201 ggccctcttg atggcggtgc cgttgtgggt gatcatccag cacaggtagg tcagctcggc
     4261 ggcacagctg gcgatcttct cgccgctggg cctctcgaat ctctccacga actgccgcac
     4321 cagcaccttg gggggggtct tgcacaggat gtcgaactgg ctcatcacct tcctcttctt
     4381 cttaggagcc atggtggcca cgccagaagc aaatgtaagc aatagatggc tctgccctga
     4441 cttttatgcc cagcccgagc tccttaagag cctccaaacc acagaagaca ggtcacaatc
     4501 tttcttccca ctgtccactg aactaagcgt acattcaaat gaggttcacg gtcttaattg
     4561 tgaattctgt ctgggacccg caggttcaag cataaaatgg gcctctctgc aaataatcac
     4621 tggcgtgtga tatttttaag aaagcaaagc cattgtttct gattgaagga aagtcagtgc
     4681 tgttttgaag agaagcacga ttaggcagtg gcgctctctt tcaccactgg gaattagacg
     4741 ccgtttgaga gatttgtgag ctccaggaca tttgccagat gtgcttgatt ctacatttca
     4801 gacaacaaag gggataatgt tggcgggaat cttgacggca ctgctgatga tcgggatatt
     4861 ctttgaggat ggtactgaga cctttcaggg gaggatttca gcttttacat aaactaattt
     4921 gggatgctca ctctctgtct tgcacaaagc aaatttagtt cattgttagc tgtatctgct
     4981 aaaatggtta caagggtttt tttccttgaa tattaataat gcgcctttca ttggagatgc
     5041 tcgaatgggg tgtcattaat tggggggggg gtagtaagga ttagtggtat agttctctcc
     5101 aaaatggtta ctactaatac catgaggacg cgtgcggccg caggaacccc tagtgatgga
     5161 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     5221 ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag
     5281 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5341 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5401 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5461 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5521 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5581 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5641 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5701 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5761 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5821 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5881 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5941 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6001 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6061 aaagtatata tgagta
//
