LOCUS       sequence-461847-        5183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0373m_3xC2 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 5183)
  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 5183)
  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: AiP12560 -
            pAAV-AiE0373m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5183
                     /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      1195..1204
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1201..1203,1204..1206,1207..1917)
                     /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(1246..1267)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1507..1526)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1854..1875
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2010..2030)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2217..2328
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2446..2586
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2446..2575
                     /label=AAV2 ITR
     rep_origin      2661..3116
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2748..2767)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2958..2979
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3133..3152)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3252..3274
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3312..3330)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3398..3502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3503..3571,3572..4363)
                     /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(3721..3740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4534..5122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5023..5042
                     /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 gcgtatgttt ggggtaaggg atggaagtgg aaaaaaaaaa
      181 aaaaaaagga aagaaaaagc aaacaaaagc caaccagcct tccagctgct gaaggagctg
      241 gggcacaacc cttgaagagt gcccctgccg ggaacagaga ggactccagc tgcctgccca
      301 ctgggccaga gcgctgcctt aaaagaactt ttgcataaaa ggaaaagagg ccaaggctcc
      361 ccttttgctg caggacacag ggccagtgtc cacaagagtt gagtgtcctg gccaaagcca
      421 aagctaggga tcattgaaaa tgcctgggca gctggcatgg ctgatgtttg gggtaaggga
      481 tggaagtgga aaaaaaaaaa aaaaaaggaa agaaaaagca aacaaaagcc aaccagcctt
      541 ccagctgctg aaggagctgg ggcacaaccc ttgaagagtg cccctgccgg gaacagagag
      601 gactccagct gcctgcccac tgggccagag cgctgcctta aaagaacttt tgcataaaag
      661 gaaaagaggc caaggctccc cttttgctgc aggacacagg gccagtgtcc acaagagttg
      721 agtgtcctgg ccaaagccaa agctagggat cattgaaaat gcctgggcag ctggcatggc
      781 tgatgtttgg ggtaagggat ggaagtggaa aaaaaaaaaa aaaaaggaaa gaaaaagcaa
      841 acaaaagcca accagccttc cagctgctga aggagctggg gcacaaccct tgaagagtgc
      901 ccctgccggg aacagagagg actccagctg cctgcccact gggccagagc gctgccttaa
      961 aagaactttt gcataaaagg aaaagaggcc aaggctcccc ttttgctgca ggacacaggg
     1021 ccagtgtcca caagagttga gtgtcctggc caaagccaaa gctagggatc attgaaaatg
     1081 cctgggcagc tggcatggct ggagctcggg ctgggcataa aagtcagggc agagccatct
     1141 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
     1201 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1261 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1321 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1381 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     1441 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1501 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1561 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1621 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
     1681 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1741 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1801 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1861 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1921 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1981 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2041 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2101 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2161 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     2221 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2281 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2341 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2401 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2461 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2521 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2581 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2641 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2701 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     2761 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2821 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2881 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2941 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     3001 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     3061 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3121 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3181 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3241 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3301 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3361 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4561 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4621 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4681 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4741 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4801 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4861 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4921 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4981 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5041 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5101 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5161 ttgctggcct tttgctcaca tgt
//
