LOCUS       sequence-461984-        5267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2100m_3xC2 is an optimized 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 5267)
  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 5267)
  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: AiP1329 - pAAV-AiE2100m_3xC2-minBG-SYFP2-WPRE-...
FEATURES             Location/Qualifiers
     source          1..5267
                     /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      1279..1288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1285..1287,1288..1290,1291..2001)
                     /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(1330..1351)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1591..1610)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1938..1959
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2094..2114)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2301..2412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2530..2670
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2530..2659
                     /label=AAV2 ITR
     rep_origin      2745..3200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2832..2851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3042..3063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3217..3236)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3336..3358
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3396..3414)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3482..3586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3587..3655,3656..4447)
                     /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(3805..3824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4618..5206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5107..5126
                     /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 gcgttaaagc aagaagggaa aatggatttc ttaatgagtc
      181 tgtatacagt gcagtatcta tgagctatgg tgtgtgtgac tgccaaggca gtcagatgct
      241 attcaagcaa ggaatgctgc tctgagatca gctagtacgt aagaaagaca tcctggctcc
      301 tggaagatga caagagtttg cctccgtagt atttgaaaga aagtggtcat ggccatcatg
      361 aatcaatcac ttcaggaaaa ggttgagcat gtgctgtgac gactgcagat ctttttatag
      421 ctgtggattc ttttatagct gctgagatac gacgatagga gtggagcggt agttatcgtt
      481 gatttgtttt aaagcaagaa gggaaaatgg atttcttaat gagtctgtat acagtgcagt
      541 atctatgagc tatggtgtgt gtgactgcca aggcagtcag atgctattca agcaaggaat
      601 gctgctctga gatcagctag tacgtaagaa agacatcctg gctcctggaa gatgacaaga
      661 gtttgcctcc gtagtatttg aaagaaagtg gtcatggcca tcatgaatca atcacttcag
      721 gaaaaggttg agcatgtgct gtgacgactg cagatctttt tatagctgtg gattctttta
      781 tagctgctga gatacgacga taggagtgga gcggtagtta tcgttgattt gttttaaagc
      841 aagaagggaa aatggatttc ttaatgagtc tgtatacagt gcagtatcta tgagctatgg
      901 tgtgtgtgac tgccaaggca gtcagatgct attcaagcaa ggaatgctgc tctgagatca
      961 gctagtacgt aagaaagaca tcctggctcc tggaagatga caagagtttg cctccgtagt
     1021 atttgaaaga aagtggtcat ggccatcatg aatcaatcac ttcaggaaaa ggttgagcat
     1081 gtgctgtgac gactgcagat ctttttatag ctgtggattc ttttatagct gctgagatac
     1141 gacgatagga gtggagcggt agttatcgtt gatttgtttc ttaaggagct cgggctgggc
     1201 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
     1261 aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1321 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1381 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     1441 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc
     1501 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1561 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1621 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1681 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1741 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1801 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1861 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     1921 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1981 ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat
     2041 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2101 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2161 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     2221 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2281 gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2341 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2401 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2461 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg
     2521 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     2581 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2641 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     2701 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     2761 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2821 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2881 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2941 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3001 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3061 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     3121 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3181 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     3241 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     3301 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     3361 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     3421 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
