LOCUS       sequence-466269-        5397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2189m 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 5397)
  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 5397)
  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: AiP1510 - pAAV-AiE2189m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      552..1140
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1041..1060
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1202..1331
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2610..2619
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2616..2618,2619..2621,2622..3332)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2661..2682)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2922..2941)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3269..3290
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3425..3445)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3632..3743
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3861..4001
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3861..3990
                     /label=AAV2 ITR
     rep_origin      4076..4531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4163..4182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4373..4394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4548..4567)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4667..4689
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4727..4745)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4813..4917
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4918..4986,4987..5397,1..381)
                     /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(5136..5155)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
       61 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      121 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      181 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      241 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      301 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      361 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      421 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      481 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
      541 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      601 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      661 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      721 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      781 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      841 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      901 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      961 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1021 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1081 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1141 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1201 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     1261 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     1321 taggggttcc tgcggccgca cgcgttgtaa gagtaataat cgccggcggc tacagttcct
     1381 gccctgctgg agtttctgcc atggcttccc tcaatgagag cctgtgattt ggatggataa
     1441 gccaaataaa attctttttc ccccttgttg cttttgacca ttatgattat cacagaaata
     1501 gaaagcaaac taggacaaat gacaagtgac gctcagaaaa actatcaaaa tcatacattg
     1561 tagaaagatg cctctgcatg aatgctgggg aaattggact tccacatgca aaagactaga
     1621 aaaaaattag atacccaatg atgtccagtt atcacactct cagatgtcag ggctcagcaa
     1681 ctgctattgc tgtgggtggt ttgataaagt tgttaaattg ttgatacttg tttgataaag
     1741 tttataaagt cactcttctt tctcagcatg tatacaaagc caaggcaggt gaaagactgg
     1801 atttaagatc cagaaccact agaagagaac aggaaaagca cttcaagacg tccctcttgg
     1861 aactgatttt ctgtgtcaca cccaaaacaa cccacaggaa accgacacaa aaagtgacaa
     1921 atgcctatga taacaaatcc aaatcaggaa gctactgcac agcaaaggga cagatgagag
     1981 agtgaagaga caccctgcat agcagaagac tgtctgccaa ccacttgtca catccagaat
     2041 atatgcagac tacctttact acagtggaga gcatgtgtgg agagcatgag tgtcaaatta
     2101 catctgaacg gtagatactc catacctaca attacctcta atttaaaccc ccaaactcct
     2161 gacaatgaca taactctgat caggcattca cactgaccca ttaaacatgt ttaggggctg
     2221 gagtcacact gaaccaagat gatggctacc tcgcaattca tgatgagagt gtgttcattt
     2281 ttttttttaa ctagcaaatt accaaataac taaaaaagag cagcgtcagg ctggaaagct
     2341 agctaagtag ctaacctgtt ttgtggtaca tgtgtgaaga gtggagtttg aattcctgga
     2401 acccacttac atctgaacca tcctcaacca tttacatttt tatccaactg ttaaagatgc
     2461 aaaggtgttc tttaaattat tttgccctac tatcagccaa tgccatctcg ccggcggagc
     2521 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
     2581 cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
     2641 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2701 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     2761 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
     2821 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2881 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2941 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3001 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3061 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     3121 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3181 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
     3241 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3301 ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa
     3361 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3421 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3481 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     3541 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3601 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     3661 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3721 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3781 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     3841 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3901 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3961 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     4021 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4081 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4141 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4201 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4261 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4321 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4381 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     4441 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4501 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4561 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4621 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4681 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4741 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4801 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4861 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4921 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4981 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5041 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5101 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5161 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5221 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5281 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5341 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgcctt
//
