LOCUS       sequence-466244-        5297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2085m 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 5297)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5297)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP1361 - pAAV-AiE2085m-minBG-SYFP2-4X2C-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      453..462
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(459..461,462..464,465..1175)
                     /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(504..525)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(765..784)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1112..1133
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1488..1508)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1695..1806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1924..2064
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1924..2053
                     /label=AAV2 ITR
     rep_origin      2139..2594
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2226..2245)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2436..2457
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2611..2630)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2730..2752
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2790..2808)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2876..2980
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2981..3049,3050..3841)
                     /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(3199..3218)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4012..4600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4501..4520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4662..4791
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ttggcctcat cattaccacc attattatta gctcattgat tgcaataatt tctaaaatta
       61 ccccttggaa taaaatatgt taagcacctg gtttatcctt tttttttttt tttaaaaaag
      121 gaacatctgc tctaaatgtg tgatagcaga ggaccaaacg tgtcttgatc tgaaggggaa
      181 gcaaaaacga tgctgacact ggggagcctt gtctgggctg gaatggatgt attgtgggat
      241 gctgagagga agagtagtca cgatcaagtt caatgctagg attacagggc tgctgagaga
      301 cggagacacg tgggaagctg agctattcca tggctactgc aggctttact tcttcttctg
      361 agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg
      421 atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
      481 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      541 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
      601 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
      661 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      721 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      781 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      841 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      901 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
      961 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1021 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     1081 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1141 ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggcctt
     1201 aaagagaccg gttcactgtg acagtaaaag agaccggttc actgtgagaa tgaaagagac
     1261 cggttcactg tgatcggaaa agagaccggt tcactgtgag cggccttgaa acccagcaga
     1321 caatgtagct cagtagaaac ccagcagaca atgtagctga atggaaaccc agcagacaat
     1381 gtagcttcgg agaaacccag cagacaatgt agctgtcgac gaattcgata tcataatcaa
     1441 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1501 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1561 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     1621 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     1681 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1741 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1801 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1861 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     1921 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     1981 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2041 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2101 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     2161 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     2221 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2281 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2341 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2401 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2461 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     2521 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2581 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2641 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2701 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2761 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2821 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2881 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2941 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3001 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3061 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3121 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3181 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3241 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3301 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3361 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3421 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3481 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3541 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3601 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3661 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3721 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3781 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3841 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3901 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3961 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4021 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4081 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4141 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4201 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4261 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4321 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4381 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4441 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4501 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4561 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4621 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     4681 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     4741 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     4801 cgcgttgact cactaaagta tttgcatgtt ttcaaagctg taaagtctgg aggtgtgctt
     4861 ttcaggagct ggggaggtta gagggaagac ctcgaacagg acacactatc actgtctttc
     4921 ttgtgccaat gcaattggaa gaaacaaaca cagccatcat ttagcccaca tgaattaggt
     4981 cctgtaaaga gtgaagggga aaaggttgtt ggaaatctaa ttagtaggct ttgcaagtgt
     5041 ggagtttatg gctacatatt cagttaaatt cagttctgtt ttgtagctat cagccaggga
     5101 agagtcaagt gccatgtgga ttgggaacat aaatagccca ttcttcatgt ccattgctct
     5161 atcattaacc ttcccactca gtcctggaaa tgaaaaaccc agcgtccttc catctgctcc
     5221 acgttattta tgatgatgcc tctaattatg ttcaattgag ttgctgtcga tgattaaagg
     5281 taataggtaa taataac
//
