LOCUS       sequence-460624-        4703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0017h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4703)
  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 4703)
  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: AiP13554 -
            pAAV-AiE0017h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4703
                     /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      715..724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(721..723,724..726,727..1437)
                     /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(766..787)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1027..1046)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1374..1395
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1530..1550)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1737..1848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1966..2106
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1966..2095
                     /label=AAV2 ITR
     rep_origin      2181..2636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2268..2287)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2478..2499
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2653..2672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2772..2794
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2832..2850)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2918..3022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3023..3091,3092..3883)
                     /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(3241..3260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4054..4642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4543..4562
                     /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 gcgtagcagg cgcccctcct ccccacctcg ctaaacaatc
      181 attgcacaaa atatgcaaat ggtataatta ctgttatttg ttttgctgat ataagtgttt
      241 gaaatgcaaa tgtcaagttt gggcgccttc atttttccaa ccctctcacc cggacatttg
      301 caaagcaggc gcccctcctc cccacctcgc taaacaatca ttgcacaaaa tatgcaaatg
      361 gtataattac tgttatttgt tttgctgata taagtgtttg aaatgcaaat gtcaagtttg
      421 ggcgccttca tttttccaac cctctcaccc ggacatttgc aaagcaggcg cccctcctcc
      481 ccacctcgct aaacaatcat tgcacaaaat atgcaaatgg tataattact gttatttgtt
      541 ttgctgatat aagtgtttga aatgcaaatg tcaagtttgg gcgccttcat ttttccaacc
      601 ctctcacccg gacatttgca agagctcggg ctgggcataa aagtcagggc agagccatct
      661 attgcttaca tttgcttctg ggatccagat ctttcgaagc tagcgctacc ggtcgccacc
      721 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      781 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      841 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      901 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      961 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1021 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1081 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1141 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
     1201 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     1261 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1321 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1381 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1441 gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     1501 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1561 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1621 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1681 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     1741 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1801 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1861 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1921 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     1981 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2041 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2101 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2161 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2221 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     2281 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2341 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2401 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2461 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     2521 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     2581 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     2641 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2701 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2761 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2821 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2881 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2941 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3001 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3061 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3121 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3181 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3241 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3301 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3361 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3421 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3481 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3541 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3601 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3661 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3721 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3781 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3841 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3901 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3961 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4021 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4081 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4141 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4201 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4261 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4321 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4381 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4441 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4501 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4561 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4621 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4681 ttgctggcct tttgctcaca tgt
//
