LOCUS       sequence-470513-        4595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0705m 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 4595)
  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 4595)
  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: AiP12712 - pAAV-AiE0705m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   32..161
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(223..811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(303..322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(982..1773,1774..1842))
                     /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     1605..1624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1843..1947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2015..2033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2071..2093)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2193..2212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2229..2684)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2366..2387)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2578..2597
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2759..2899
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2770..2899
                     /label=AAV2 ITR
     polyA_signal    3017..3128
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3315..3335
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3428..4138,4139..4141,4142..4144))
                     /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(3470..3491)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3819..3838
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4078..4099
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4141..4150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ttttaaccct tttctgtacg cgtgcggccg caggaacccc tagtgatgga gttggccact
       61 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
      121 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca
      181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      661 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
      841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
      961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     1981 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2041 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2101 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2161 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     2221 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2281 cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata
     2341 gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2401 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2461 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2521 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2581 gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt
     2641 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     2701 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcccc
     2761 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     2821 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     2881 tccatcacta ggggttcctg cggccgctcg gtccgcacgt gagatctcat gcctgctatt
     2941 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
     3001 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
     3061 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
     3121 aggcacagtc gaagatctct cgagccacgg aattgtcagt gcccaacagc cgagcccctg
     3181 tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaagaact aaccaggatt
     3241 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     3301 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     3361 cacaaatttt gtaatccaga ggttgattat gatatcgaat tcgcggccgc ggcgcgccgt
     3421 cgacttactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     3481 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tggtagctca
     3541 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     3601 ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg gatcttgaag ttggccttga
     3661 tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag ttgtactcca
     3721 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     3781 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     3841 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     3901 gctgcttcat gtggtcgggg tagcgggcga agcactgcac gccgtagccc agggtggtca
     3961 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatcagcttc agggtcagct
     4021 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     4081 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     4141 ccatggtggc gaccggtagc gctagcttcg aaagatctgg atcccagaag caaatgtaag
     4201 caatagatgg ctctgccctg acttttatgc ccagcccgag ctctgatggc caggggaaag
     4261 aatacattgt gggccactcc ttttaaaaac atttaattac aggagggaag gaggaagaaa
     4321 aagaaaaaaa cttcgtactc ttcattttgt tgtatctaac gaatttttaa agtaatttta
     4381 aatctttgac acacatccag aggcaccagt tttggtagat gttaaagtgt caagtgatta
     4441 ggcaaaaaag caaatcaatt ccattcttta aacatccagt ttgtaattag tttttcttgg
     4501 atggtttcca gagaagatac attgctctaa tttcaatatt ttgcgacagt ttgcaaagat
     4561 gatggttaaa aaatgtagtt actaacattc tacat
//
