LOCUS       sequence-449786-        5113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0526h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical Sst+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5113)
  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 5113)
  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: AiP13596 -
            pAAV-AiE0526h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      1125..1134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1131..1133,1134..1136,1137..1847)
                     /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(1176..1197)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1437..1456)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1784..1805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1940..1960)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2147..2258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2376..2516
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2376..2505
                     /label=AAV2 ITR
     rep_origin      2591..3046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2678..2697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2888..2909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3063..3082)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3182..3204
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3242..3260)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3328..3432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3433..3501,3502..4293)
                     /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(3651..3670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4464..5052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4953..4972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtactaa gatgtttatg gcaatcgccc
      181 tgcaagtgtc tatttaattc ctgaaaatgt catttactgg tgtgtttctg taaaaatcat
      241 ttctgaaaag ttttgtcagc ttggcttagc agctggggga aaaaagcagc aaaatgagtc
      301 ccctcaatca agcctgtgat tttttatttt ttcagactca tgaatcctcc tgagaattag
      361 gcttatgaca agcacccacc ccctccacct actccctggt ttaaaccagg tgcctgctga
      421 cacgaaggaa gattccgggg cattttgact aagatgttta tggcaatcgc cctgcaagtg
      481 tctatttaat tcctgaaaat gtcatttact ggtgtgtttc tgtaaaaatc atttctgaaa
      541 agttttgtca gcttggctta gcagctgggg gaaaaaagca gcaaaatgag tcccctcaat
      601 caagcctgtg attttttatt ttttcagact catgaatcct cctgagaatt aggcttatga
      661 caagcaccca ccccctccac ctactccctg gtttaaacca ggtgcctgct gacacgaagg
      721 aagattccgg ggcattttga ctaagatgtt tatggcaatc gccctgcaag tgtctattta
      781 attcctgaaa atgtcattta ctggtgtgtt tctgtaaaaa tcatttctga aaagttttgt
      841 cagcttggct tagcagctgg gggaaaaaag cagcaaaatg agtcccctca atcaagcctg
      901 tgatttttta ttttttcaga ctcatgaatc ctcctgagaa ttaggcttat gacaagcacc
      961 caccccctcc acctactccc tggtttaaac caggtgcctg ctgacacgaa ggaagattcc
     1021 ggggcatttt ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca
     1081 tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca
     1141 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1201 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1261 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1321 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1381 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1441 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1501 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1561 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1621 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1681 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1741 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1801 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg
     1861 cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat
     1921 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     1981 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2041 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2101 ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt
     2161 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2221 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2281 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2341 aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     2401 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2461 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     2521 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     2581 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2641 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2701 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2761 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2821 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2881 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct
     2941 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3001 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     3061 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3121 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3181 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3241 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3301 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3361 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3421 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3481 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3541 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3601 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3661 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3721 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3781 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3841 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3901 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3961 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4021 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4081 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4141 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4201 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4261 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4321 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4381 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4441 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4501 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4561 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4621 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4681 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4741 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4801 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4861 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4921 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4981 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5041 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5101 tttgctcaca tgt
//
