LOCUS       sequence-461929-        5471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2118m_3xC3 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 5471)
  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 5471)
  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: AiP14481 -
            pAAV-AiE2118m_3xC3-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5471
                     /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      1483..1492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1489..1491,1492..1494,1495..2205)
                     /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(1534..1555)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1795..1814)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2142..2163
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2298..2318)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2505..2616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2734..2874
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2734..2863
                     /label=AAV2 ITR
     rep_origin      2949..3404
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3036..3055)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3246..3267
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3421..3440)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3540..3562
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3600..3618)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3686..3790
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3791..3859,3860..4651)
                     /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(4009..4028)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4822..5410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5311..5330
                     /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 gcgtctgctc attaaacaaa tctctcaggc ggacttaacg
      181 tgaattagca cttctctgaa aggggcagcc ttgatgagca gcagacagct cagatcattt
      241 ggaaggtcac agtttttttt ttattattat ataagcccag ttgttattaa tgtattcata
      301 tttgaagaaa ctgagcagag acctgctttc attcaatcta gtgagcaaaa cagtacgttt
      361 tcctaactgg aaaacaaaac ataaatgcta atcaggtccc agcagagatc aaatcaagct
      421 aaatatagca ggcacccttt gtggtttttt ttgttttgtt ttgttttgtt ttttgttttt
      481 ttttaataca gaaattaagg aaagaaaaaa aaccattgaa ttaattgtct tcctctgtgc
      541 ttcgcacctg acaccagcgc tgctcattaa acaaatctct caggcggact taacgtgaat
      601 tagcacttct ctgaaagggg cagccttgat gagcagcaga cagctcagat catttggaag
      661 gtcacagttt tttttttatt attatataag cccagttgtt attaatgtat tcatatttga
      721 agaaactgag cagagacctg ctttcattca atctagtgag caaaacagta cgttttccta
      781 actggaaaac aaaacataaa tgctaatcag gtcccagcag agatcaaatc aagctaaata
      841 tagcaggcac cctttgtggt tttttttgtt ttgttttgtt ttgttttttg ttttttttta
      901 atacagaaat taaggaaaga aaaaaaacca ttgaattaat tgtcttcctc tgtgcttcgc
      961 acctgacacc agcgctgctc attaaacaaa tctctcaggc ggacttaacg tgaattagca
     1021 cttctctgaa aggggcagcc ttgatgagca gcagacagct cagatcattt ggaaggtcac
     1081 agtttttttt ttattattat ataagcccag ttgttattaa tgtattcata tttgaagaaa
     1141 ctgagcagag acctgctttc attcaatcta gtgagcaaaa cagtacgttt tcctaactgg
     1201 aaaacaaaac ataaatgcta atcaggtccc agcagagatc aaatcaagct aaatatagca
     1261 ggcacccttt gtggtttttt ttgttttgtt ttgttttgtt ttttgttttt ttttaataca
     1321 gaaattaagg aaagaaaaaa aaccattgaa ttaattgtct tcctctgtgc ttcgcacctg
     1381 acaccagcgg agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     1441 tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag
     1501 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1561 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1621 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1681 ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     1741 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1801 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1861 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1921 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     1981 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     2041 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     2101 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2161 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg
     2221 ccgcggccgc gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg
     2281 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2341 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2401 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2461 cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg
     2521 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2581 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2641 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2701 gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     2761 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     2821 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     2941 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3001 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     3061 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3121 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     3181 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3241 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     3301 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     3361 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     3421 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     3481 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3541 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     3601 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3661 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3721 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3781 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3841 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3901 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3961 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4021 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4081 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4141 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4201 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4261 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4321 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4381 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4441 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4501 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4561 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4621 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4681 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4741 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4801 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4861 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4921 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4981 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5041 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5101 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5161 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5221 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5281 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5341 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5401 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5461 tgctcacatg t
//
