LOCUS       sequence-466291-        4872 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2364m 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 4872)
  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 4872)
  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: AiP1689 - pAAV-AiE2364m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(34..138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     206..224
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(262..284)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     384..403
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(420..875)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(557..578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     769..788
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   950..1090
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   961..1090
                     /label=AAV2 ITR
     polyA_signal    1208..1319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1506..1526
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1619..2329,2330..2332,2333..2335))
                     /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(1661..1682)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2010..2029
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2269..2290
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2332..2341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3095..3224
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3286..3874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3366..3385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4045..4836,4837..4872,1..33))
                     /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     4668..4687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
       61 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
      121 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
      181 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
      241 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
      301 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      361 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
      421 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
      481 ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat
      541 agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      601 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
      661 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
      721 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
      781 gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
      841 acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg
      901 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag
      961 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
     1021 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     1081 aggggttcct gcggccgctc ggtccgcacg tgagatctca tgcctgctat tgtcttccca
     1141 atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag
     1201 acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag
     1261 ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt
     1321 cgaagatctc tcgagccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc
     1381 gggcaaggca ggcggcgatg agttccgccg tggcaagaac taaccaggat ttatacaagg
     1441 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1501 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1561 tgtaatccag aggttgatta tgatatcgaa ttcgcggccg cggcgcgccg tcgacttact
     1621 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     1681 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctggtagctc aggtagtggt
     1741 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     1801 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct
     1861 tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     1921 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     1981 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     2041 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     2101 tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc cagggtggtc acgagggtgg
     2161 gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc ttgccgtagg
     2221 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     2281 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     2341 cgaccggtag cgctagcttc gaaagatctg gatcccagaa gcaaatgtaa gcaatagatg
     2401 gctctgccct gacttttatg cccagcccga gctccgatga gacaaagagg ggaaagtgac
     2461 ccagagaagg ggccccctat aaggggaggg aagccaaact gtggctgcaa attgagaaaa
     2521 cattgacaca gagatggaga gttttggagt tttggaagga gaaaggctaa ctggcttttg
     2581 agatgtggaa tcggaaaggg aaagacatca cgccatggga attttggaaa acaagaatca
     2641 caaagctact tacaactgat agagtgtgga tgcagaagac aagagggaac actacaggtg
     2701 cgtaggaagg aggctttgaa acatgaacat caaggcctgt ccaagtcagt gaagatgtgc
     2761 ccgcctggca gcttcctccg aatgtgctaa ttacactttg tgttcttcaa caaaaatagt
     2821 ttctttgtga ggagggctgg agtccaaggg tgggagagag catcttctca caggcagcca
     2881 gtaggcagcc ccacttgaag gataataaat atattgttat gcaacagcac actcacgtct
     2941 catgaaggct ggctcttact atcctggtga ggatgagcat tctgagggaa aaagagaagg
     3001 tggcagtacg gctcagtgat gaggtccctc tttgtatcat ttatgccagg ctacagtagc
     3061 agccgattat tactcttaca acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc
     3121 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3181 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg
     3241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3721 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3961 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4021 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4081 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4141 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4201 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4261 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4321 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4381 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4441 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4501 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4561 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4621 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4681 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4741 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4801 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4861 cgcaaaaaag gg
//
