LOCUS       sequence-470554-        4591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0585m 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 4591)
  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 4591)
  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: AiP12874 - pAAV-AiE0585m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   30..159
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(221..809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(301..320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(980..1771,1772..1840))
                     /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     1603..1622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1841..1945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2013..2031
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2069..2091)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2191..2210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2227..2682)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2364..2385)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2576..2595
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2757..2897
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2768..2897
                     /label=AAV2 ITR
     polyA_signal    3015..3126
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3313..3333
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3426..4136,4137..4139,4140..4142))
                     /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(3468..3489)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3817..3836
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4076..4097
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4139..4148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tgaaagctaa ccagtacgcg tgcggccgca ggaaccccta gtgatggagt tggccactcc
       61 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
      121 cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc
      181 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      241 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      301 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      361 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      421 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      481 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      541 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      601 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
      661 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      721 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      781 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      841 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      901 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
      961 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1021 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1081 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1141 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1201 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1261 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1321 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1381 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1441 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1501 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1561 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1621 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1681 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1741 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1801 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1861 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1921 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     1981 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     2041 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     2101 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     2161 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     2221 accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2281 gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc
     2341 ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2401 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2461 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2521 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2581 agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2641 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg
     2701 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg
     2761 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
     2821 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     2881 catcactagg ggttcctgcg gccgctcggt ccgcacgtga gatctcatgc ctgctattgt
     2941 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
     3001 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
     3061 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
     3121 gcacagtcga agatctctcg agccacggaa ttgtcagtgc ccaacagccg agcccctgtc
     3181 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caagaactaa ccaggattta
     3241 tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa
     3301 gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca
     3361 caaattttgt aatccagagg ttgattatga tatcgaattc gcggccgcgg cgcgccgtcg
     3421 acttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     3481 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg gtagctcagg
     3541 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     3601 tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg
     3661 ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc
     3721 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     3781 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     3841 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     3901 tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc cgtagcccag ggtggtcacg
     3961 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg
     4021 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     4081 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     4141 atggtggcga ccggtagcgc tagcttcgaa agatctggat cccagaagca aatgtaagca
     4201 atagatggct ctgccctgac ttttatgccc agcccgagct cttgctttag tctcaagatc
     4261 ccaatactat gttattttcc actccccctg ccaccgtacg gtcttcactg tcctgcaggg
     4321 tcttgtctgt agcatatatt aaagaatact ttcaaaatga acaaaactga ttgtacatat
     4381 tgtgtgtcat acatcaagac aagaatagat ttggctctgt gtcctagctg ctgtgacaga
     4441 tgtcctggca tctggtagtg tgtcctttgt acagagctga cagtcattag ctggtgacca
     4501 cacagaaagc taagaatccc agaagggtct ctataaatca gtccagtaca tcacaaaaga
     4561 aacactggtt gactgacttt aactgtcaga t
//
