LOCUS       sequence-461339-        5248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0078h_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 5248)
  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 5248)
  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: AiP14803 -
            pAAV-AiE0078h_3xC2-minBG-SYFP2-10aa...
FEATURES             Location/Qualifiers
     source          1..5248
                     /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      874..883
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(880..882,883..885,886..1596)
                     /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(925..946)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1186..1205)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1533..1554
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1627..2004
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2097..2117)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2304..2415
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2726..3181
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2813..2832)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3023..3044
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3198..3217)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3317..3339
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3377..3395)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3463..3567
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3568..3636,3637..4428)
                     /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(3786..3805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4599..5187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5088..5107
                     /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 gcgtattagc aatttggcag gtacccgagg gccataccta
      181 atctgcataa aatgaagcag attgcaaccg ccctcatctt ttttattttt aaactggttt
      241 ttgaagcaga gcataaaatc tcagagggag agacagaaga tgctagtgca tacattttcc
      301 ttcatgcctt tattttcatt ctttttgcac aaaccatctt cctgaatggc tgtttaatta
      361 gcaatttggc aggtacccga gggccatacc taatctgcat aaaatgaagc agattgcaac
      421 cgccctcatc ttttttattt ttaaactggt ttttgaagca gagcataaaa tctcagaggg
      481 agagacagaa gatgctagtg catacatttt ccttcatgcc tttattttca ttctttttgc
      541 acaaaccatc ttcctgaatg gctgtttaat tagcaatttg gcaggtaccc gagggccata
      601 cctaatctgc ataaaatgaa gcagattgca accgccctca tcttttttat ttttaaactg
      661 gtttttgaag cagagcataa aatctcagag ggagagacag aagatgctag tgcatacatt
      721 ttccttcatg cctttatttt cattcttttt gcacaaacca tcttcctgaa tggctgttta
      781 gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg
      841 gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
      901 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      961 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     1021 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     1081 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1141 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1201 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1261 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1321 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1381 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1441 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1501 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1561 gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc tggaggctcc
     1621 ggaggcccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa gaaggcggtg
     1681 actaaggcgc agaagaaagg cggcaagaag cgcaagcgca gccgcaagga gagctattcc
     1741 atctatgtgt ataaggttct gaagcaggtc caccctgaca ccggcatttc gtccaaggcc
     1801 atgggcatca tgaactcgtt tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc
     1861 cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca gacggccgtg
     1921 cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac taaggccatc
     1981 accaagtaca ccagcgctaa gtaatgagtc gacggcgcgc cgcggccgcg aattcgatat
     2041 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2101 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2161 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     2221 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2281 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2341 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2401 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2461 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     2521 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2581 cgctcactga ggccgggcga ccaaaggtcg cccggcctca gtgagcgagc gagcgcgcag
     2641 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2701 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2761 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     2821 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2881 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2941 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3001 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3061 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     3121 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     3181 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3241 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3301 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3361 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3421 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3481 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3541 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3601 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3661 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3721 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3781 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3841 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3901 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3961 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4021 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4081 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4141 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4201 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4261 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4321 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4381 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4441 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4501 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4561 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4621 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4681 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4741 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4801 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4861 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4921 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4981 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5041 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5101 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5161 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5221 gccttttgct ggccttttgc tcacatgt
//
