LOCUS       sequence-470590-        4931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0389m 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 4931)
  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 4931)
  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: AiP13061 - pAAV-AiE0389m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(36..55)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     246..267
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(421..440)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     540..562
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(600..618)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        686..790
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(791..859,860..1651)
                     /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(1009..1028)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1822..2410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2311..2330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2472..2601
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3414..3423
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3420..3422,3423..3425,3426..4136)
                     /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(3465..3486)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3726..3745)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4073..4094
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4229..4249)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4436..4547
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4665..4805
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4665..4794
                     /label=AAV2 ITR
     rep_origin      join(4880..4931,1..404)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
       61 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      121 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
      181 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      241 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
      301 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
      361 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
      421 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
      481 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
      541 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
      601 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
      661 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      721 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      781 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      841 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      901 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      961 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1021 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1081 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1141 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1201 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1261 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1321 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1381 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1441 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1501 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1561 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1621 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1681 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1741 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1801 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1861 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1921 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1981 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2041 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2101 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2161 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2221 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2281 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2341 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2401 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2461 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
     2521 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     2581 actccatcac taggggttcc tgcggccgca cgcgttgccc agaagctatg tgaccacatg
     2641 taagtctccc attgcatgga gatagaaaat ttagagctag tagccctgca gaatctagtt
     2701 gcagacccct tcatgggccc cgaatttcct gtcttgtcat cacagcaaag cgaggactgg
     2761 gaatggtctc agcatagtgc tggctcagag ccggcaaagt tcctgctgtt cttcaaggga
     2821 aagttgtttg ggttctttgt ccagtaactt ggtaacaaga aaactctcac gctggtcaaa
     2881 aaaaaaaaaa aaaggaaaag cctcgggacc cccatgccac tcccagcttc ctctgctccc
     2941 cccttgaaag cagagactag agcagtgggt ggtattgggc ctgttctgcc cagaggctga
     3001 ggagggtggc ccgcccccaa ctgccagcca gtacaagaca aaagaccctt gacagttccc
     3061 tggggcggtt cagctgtggc ccggctggac aaagcaaggg tacagggctc ggtacagctc
     3121 tgcaaagtcc ttggggcttg tcttggcgga gacttgggct tttgagaagc tctgtccagg
     3181 gaagctctat aaacccagac aacaaaatga cccctctcct cttacttgaa gccagaagtc
     3241 aaatggaagc aacaggccct ggcctcaccc tctttcaccc caaggccaca tctctgaact
     3301 agaggtggca gaagcacctt gagctcgggc tgggcataaa agtcagggca gagccatcta
     3361 ttgcttacat ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca
     3421 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3481 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3541 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3601 tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
     3661 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3721 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3781 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3841 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     3901 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     3961 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4021 acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4081 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     4141 tcgacggcgc gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt
     4201 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4261 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4321 ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4381 ggacaggggc tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg
     4441 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4501 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4561 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4621 gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     4681 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4741 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     4801 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4861 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4921 gttacgcgca g
//
