LOCUS       sequence-461906-        5063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0226h_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 5063)
  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 5063)
  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: AiP12568 -
            pAAV-AiE0226h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5063
                     /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      1075..1084
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1081..1083,1084..1086,1087..1797)
                     /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(1126..1147)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1387..1406)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1734..1755
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1890..1910)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2097..2208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2326..2466
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2326..2455
                     /label=AAV2 ITR
     rep_origin      2541..2996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2628..2647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2838..2859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3013..3032)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3132..3154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3192..3210)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3278..3382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3383..3451,3452..4243)
                     /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(3601..3620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4414..5002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4903..4922
                     /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 gcgtccactt ttgtatttac cattattgca aaatcaatca
      181 aatgtgagca aaagcaaagg caatttgaat gactcaaaat tgaaatttga gctgccaaat
      241 caagcagtaa aacaattttt accagctcta ttgattgtta gaaagataag ttataaactt
      301 tatttcaagt aaatttctaa aagatctggg aatgtgatta ttccaaggca gatggcgaag
      361 acctttattt ccactgatta ttcacagatg caaattatta tgcaactgga agcatactaa
      421 gatccacttt tgtatttacc attattgcaa aatcaatcaa atgtgagcaa aagcaaaggc
      481 aatttgaatg actcaaaatt gaaatttgag ctgccaaatc aagcagtaaa acaattttta
      541 ccagctctat tgattgttag aaagataagt tataaacttt atttcaagta aatttctaaa
      601 agatctggga atgtgattat tccaaggcag atggcgaaga cctttatttc cactgattat
      661 tcacagatgc aaattattat gcaactggaa gcatactaag atccactttt gtatttacca
      721 ttattgcaaa atcaatcaaa tgtgagcaaa agcaaaggca atttgaatga ctcaaaattg
      781 aaatttgagc tgccaaatca agcagtaaaa caatttttac cagctctatt gattgttaga
      841 aagataagtt ataaacttta tttcaagtaa atttctaaaa gatctgggaa tgtgattatt
      901 ccaaggcaga tggcgaagac ctttatttcc actgattatt cacagatgca aattattatg
      961 caactggaag catactaaga tgagctcggg ctgggcataa aagtcagggc agagccatct
     1021 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
     1081 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1141 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1201 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1261 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     1321 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1381 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1441 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1501 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
     1561 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1621 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1681 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1741 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1801 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1861 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1921 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1981 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2041 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     2101 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2161 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2221 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2281 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2341 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2401 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2461 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2521 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2581 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     2641 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2701 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2761 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2821 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     2881 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     2941 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3001 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3061 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3121 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3181 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3241 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3301 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3361 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3421 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3481 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3541 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3601 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3661 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3721 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3781 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3841 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3901 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3961 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4021 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4081 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4141 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4201 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4261 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4321 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4381 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4441 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4501 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4561 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4621 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4681 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4741 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4801 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4861 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4921 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4981 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5041 ttgctggcct tttgctcaca tgt
//
