LOCUS       sequence-466329-        5197 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2417m 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 5197)
  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.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: AiP1914 - pAAV-AiE2417m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      493..502
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(499..501,502..504,505..1215)
                     /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(544..565)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(805..824)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1152..1173
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1308..1328)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1515..1626
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1744..1884
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1744..1873
                     /label=AAV2 ITR
     rep_origin      1959..2414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2046..2065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2256..2277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2431..2450)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2550..2572
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2610..2628)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2696..2800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2801..2869,2870..3661)
                     /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(3019..3038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3832..4420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4321..4340
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4482..4611
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ccaccccatg aaaggactgt gccatttgcc aatgacactc agcacacagc agccacagct
       61 ttgctgcttc tgtccctaca ttgggactcc cagctgggac acatggcaga cagtgcaatc
      121 tgagtggtgt gagggaccta aagttattgg tgtatgacag ctttcccttg agatgggaaa
      181 cacacaggcc tgtaatccca gcagctgaga ggctgaagca ggaggaccat gagtttgagg
      241 ccaacccgag ctgtatagtg acagtctgtc tccaaaaggg ctaaggggga actggagact
      301 gttcggttca gtgcaggcct tataagcctg tagggctgat tttaatctct tggaacctgt
      361 ataaagaaat atgcatactc tcagcgctag caaaaaacgg agctcgggct gggcataaaa
      421 gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta
      481 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      541 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      601 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
      661 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc
      721 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      781 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      841 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      901 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
      961 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     1021 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1081 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
     1141 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1201 gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata tcataatcaa
     1261 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1321 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1381 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     1441 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     1501 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1561 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1621 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1681 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     1741 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     1801 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     1861 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     1921 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     1981 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     2041 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2101 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2161 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2221 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2281 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     2341 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2401 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2461 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2521 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2581 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2641 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2701 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2761 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2821 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2881 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2941 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3001 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3061 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3121 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3181 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3241 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3301 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3361 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3421 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3481 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3541 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3601 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3661 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3721 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3781 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3841 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3901 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3961 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4021 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4081 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4141 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4201 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4261 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4321 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4381 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4441 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     4501 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     4561 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     4621 gccaaggctt tacagaaagc aagatccagg actggggttt gaactgggat tctctctgat
     4681 agctaacact ctcacttcta cctgaataac acaaataatc ccaggcaggt ccagtgtgca
     4741 gcaaaggctg aactgacacc aacggagccc agagcacttt agcaataaag ctgtaaatga
     4801 aaagaaatgc attcaagctg ttaaccttat agagatggaa aattaattat cataataatg
     4861 gcttccacac ttctatccaa tgctggcttc tggaggctct actggagaca gaattcactc
     4921 ttgtgtttat ttattcactc atccctttgg tggctccctg cagcacctgg tctgtgtatg
     4981 tcaggagtca ggctcactgg gaaggatgag aaggagaagc agacaagatg cagagtgaaa
     5041 aggtgttgct accacacaat gtcctttccc agcctgtcat tcattctaaa ctaccagtcc
     5101 ctgaaacaaa ctcaatgagg ctctagacgg gcttgtatac cctccgtcta gacgggtgtc
     5161 caattgagcg ccattatccc tacagacacg cccttcc
//
