LOCUS       sequence-473567-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0658m 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 4733)
  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 4733)
  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: AiP12752 - pAAV-AiE0658m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      745..754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(751..753,754..756,757..1467)
                     /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(796..817)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1057..1076)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1404..1425
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1560..1580)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1767..1878
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1996..2136
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1996..2125
                     /label=AAV2 ITR
     rep_origin      2211..2666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2298..2317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2508..2529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2683..2702)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2802..2824
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2862..2880)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2948..3052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3053..3121,3122..3913)
                     /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(3271..3290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4084..4672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4573..4592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctgcgc caccagctgc cgcagaggcg tctcaacggc
      181 tccgagctgc aaccgccacc gccaccgcca ccgccgccga tgtcgctacg ccgctcccgc
      241 gcggctcccg ctcggcgccc gctagcgctg ggccgtacca agtgctcgcg caggcacggg
      301 tggcgctaca gccgccgagc gtcgtggctc gggagctgag ggcctgagtc ccctagagcc
      361 cacaggtgcc cctggttgcg accaattgca agcagatgcc gtgggagggg agggagaggg
      421 gaggcccgga gagaggcacg caggagctgt ccctctcctc cacccccccc acggctggca
      481 cagtggtcgc gcccggcccg cccctgctgg ctgctctgcg tgtgtgtatt tgtatgtgta
      541 ctgtatgtaa aagacgggga gggaggaaac gattgttgtg gacccggttt gggggcgggt
      601 atccgggcct gtagcccacg gctgcatgtt taagagaatg tggcagccca agagctcggg
      661 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      721 ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
      781 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      841 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
      901 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
      961 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1021 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1081 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1141 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1201 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1261 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1321 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1381 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1441 actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     1501 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1561 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1621 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     1681 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1741 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     1801 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1861 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1921 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     1981 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2041 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2101 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2161 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2221 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2281 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2341 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2401 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     2461 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2521 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     2581 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2641 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     2701 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2761 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     2821 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     2881 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     2941 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3001 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3061 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3121 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3181 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3241 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3301 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3361 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3421 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3481 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3541 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3601 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3661 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3721 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3781 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3841 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3901 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3961 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4021 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4081 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4141 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4201 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4261 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4321 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4381 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4441 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4501 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4561 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4621 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4681 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
