LOCUS       sequence-470585-        4602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0679m 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 4602)
  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 4602)
  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: AiP12945 - pAAV-AiE0679m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(85..104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      898..1486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1387..1406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1548..1677
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2161..2170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2167..2169,2170..2172,2173..2883)
                     /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(2212..2233)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2473..2492)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2820..2841
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2976..2996)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3183..3294
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3412..3552
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3412..3541
                     /label=AAV2 ITR
     rep_origin      3627..4082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3714..3733)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3924..3945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4099..4118)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4218..4240
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4278..4296)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4364..4468
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4469..4537,4538..4602,1..727)
                     /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"
ORIGIN
        1 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
       61 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      121 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      181 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      241 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
      301 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
      361 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
      421 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
      481 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
      541 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
      601 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
      661 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
      721 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
      781 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
      841 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
      901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
      961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     1321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     1441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     1501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
     1561 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
     1621 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     1681 gccgcacgcg ttccatttga tcatcattca tggcatacac aaatataaaa tttttcaaga
     1741 aaataaaatc taaaaaccaa aaagaagtct ccaaagagaa tagcctgaaa aggaaacttc
     1801 aacaatatcc cacccatcac atctaccagc atatttttac acagacatta gtcatttggt
     1861 cactagaact aaaagacagg ataaaggaac tgttctttta gtcatatggc aaccatccca
     1921 gattataatt agagattcct tttggtgaac taaggaaaac ataaatgtaa agctagaagc
     1981 tgtccatgta gtgcttgctt atttttttac ttaaattcat ttcctcaaaa attgtttcaa
     2041 tcttattttg tccttaaatt tctataggag ctcgggctgg gcataaaagt cagggcagag
     2101 ccatctattg cttacatttg cttctgggat ccagatcttt cgaagctagc gctaccggtc
     2161 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2221 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2281 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     2341 cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac
     2401 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2461 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2521 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2581 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     2641 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     2701 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2761 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     2821 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2881 aagtaagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac
     2941 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3001 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3061 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     3121 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc
     3181 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3241 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3301 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3361 ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc
     3421 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3481 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3541 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3601 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3661 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     3721 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3781 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3841 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3901 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3961 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     4021 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     4081 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4141 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4201 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4261 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4321 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4381 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4441 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4501 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4561 aaaagatgct gaagatcagt tgggtgcacg agtgggttac at
//
