LOCUS       sequence-461859-        4637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0729m 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 4637)
  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 4637)
  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: AiP12756 - pAAV-AiE0729m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4637
                     /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      649..658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(655..657,658..660,661..1371)
                     /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(700..721)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(961..980)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1308..1329
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1464..1484)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1671..1782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1900..2040
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1900..2029
                     /label=AAV2 ITR
     rep_origin      2115..2570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2202..2221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2412..2433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2587..2606)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2706..2728
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2766..2784)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2852..2956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2957..3025,3026..3817)
                     /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(3175..3194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3988..4576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4477..4496
                     /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 gcgttgttct tttccagagg cccagtggag tctgaggcct
      181 ttatcctggg attaatattt caatacatat aacagaatgt acgcatttgc aaaggaagta
      241 agtcacattt aaatatgggt atcagaacag ttaaaattta aatatagtaa tacatgtttc
      301 ttgcattggc tttataaaat taacttttgg ttggcataca aagatgtgag tttcatgtgc
      361 catttttatg catatgtatc atcttgtcta gttctgtttg ctcctccctt cccctgcctc
      421 ctttgtatca gaggagacaa gggtgcccac ttccttcact cttactcaag atagtactta
      481 atattaacta gagcagtaag acatgagaaa aaactaaaaa ggataaaaat aacaaaggag
      541 agtcaaatta tccctgagct cgggctgggc ataaaagtca gggcagagcc atctattgct
      601 tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg
      661 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      721 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      781 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      841 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      901 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      961 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1021 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1081 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1141 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     1201 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1261 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1321 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac
     1381 ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa
     1441 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     1501 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     1561 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     1621 ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc
     1681 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1741 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1801 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1861 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     1921 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     1981 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     2041 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2101 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2161 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     2221 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2281 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2341 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2401 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     2461 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     2521 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2581 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2641 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2701 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2761 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2821 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2881 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2941 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3001 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3061 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3121 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3181 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3241 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3301 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3361 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3421 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3481 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3541 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3601 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3661 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3721 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3781 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3841 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3901 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3961 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4021 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4081 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4141 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4201 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4261 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4321 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4381 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4441 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4501 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4561 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4621 gccttttgct cacatgt
//
