LOCUS       sequence-470543-        4652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0727m 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 4652)
  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 4652)
  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: AiP12866 - pAAV-AiE0727m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     279..298
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   440..569
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1103..1112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1109..1111,1112..1114,1115..1825)
                     /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(1154..1175)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1415..1434)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1762..1783
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1918..1938)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2125..2236
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2354..2494
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2354..2483
                     /label=AAV2 ITR
     rep_origin      2569..3024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2656..2675)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2866..2887
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3041..3060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3160..3182
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3220..3238)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3306..3410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3411..3479,3480..4271)
                     /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(3629..3648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4442..4652,1..378)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
       61 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      421 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
      481 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      541 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtagagaat atggtcctct
      601 gaaaacatca gagtgtccta aatactttga gcctggatca gctaatctcc aacaagaacc
      661 tggaccagaa tttgtcagtt tgtcagcttt tctttcctgt gtgagaaaca gacagttgct
      721 gatcagaata gcaaaggctc ctgttgtgtc ttcaaatgta tgcttcatat agcttaagaa
      781 ttggtaacag tcccagatac acaaatcatt ttgatctttc tagttgcata tgactatatt
      841 tatgtatcca ataaatatgg ctatctgaga attgcaattc caaatgtttg caagggctgg
      901 cagattacct gagtttgaaa tgggtcatgt gtgaggaatg gctgggtctc tcaacctaag
      961 tgctgttggt attaggggcc aggtacattt ggctgttgga agtgatgctg agctcgggct
     1021 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
     1081 ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1141 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1201 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     1261 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
     1321 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1381 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1441 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1501 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1561 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1621 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1681 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1741 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1801 tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc gaattcgata
     1861 tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1921 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1981 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     2041 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2101 ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2161 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2221 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2281 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
     2341 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2401 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2461 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2521 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2581 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2641 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2701 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2761 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2821 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2881 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     2941 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3001 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3061 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3121 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3181 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3241 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3301 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3361 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3421 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3481 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3541 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3601 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3661 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3721 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3781 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3841 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3901 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3961 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4021 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4081 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4141 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4201 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4261 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4321 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4381 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4441 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4501 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4561 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4621 tcaagaactc tgtagcaccg cctacatacc tc
//
