LOCUS       sequence-460602-        4928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0528h_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 4928)
  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 4928)
  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: AiP13599 -
            pAAV-AiE0528h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4928
                     /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      940..949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(946..948,949..951,952..1662)
                     /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(991..1012)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1252..1271)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1599..1620
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1755..1775)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1962..2073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2191..2331
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2191..2320
                     /label=AAV2 ITR
     rep_origin      2406..2861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2493..2512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2703..2724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2878..2897)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2997..3019
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3057..3075)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3143..3247
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3248..3316,3317..4108)
                     /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(3466..3485)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4279..4867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4768..4787
                     /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 gcgtcattgc agcaatgctg tcaaagtctg gatccctgac
      181 tggtggctgc aaactgttta ttaacagctt gatgctcatt aaggacattg agccagaata
      241 taagtcaact gtgtcactaa gtgtgctatt tagcatagct ggcattatgt gtgttgtggg
      301 agaggcaaga ctctcaatga aagaggcaat acattgattt tcattctgat gtaagccatt
      361 tcacttttga ttgggactca ttgcagcaat gctgtcaaag tctggatccc tgactggtgg
      421 ctgcaaactg tttattaaca gcttgatgct cattaaggac attgagccag aatataagtc
      481 aactgtgtca ctaagtgtgc tatttagcat agctggcatt atgtgtgttg tgggagaggc
      541 aagactctca atgaaagagg caatacattg attttcattc tgatgtaagc catttcactt
      601 ttgattggga ctcattgcag caatgctgtc aaagtctgga tccctgactg gtggctgcaa
      661 actgtttatt aacagcttga tgctcattaa ggacattgag ccagaatata agtcaactgt
      721 gtcactaagt gtgctattta gcatagctgg cattatgtgt gttgtgggag aggcaagact
      781 ctcaatgaaa gaggcaatac attgattttc attctgatgt aagccatttc acttttgatt
      841 gggactgagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
      901 ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc
      961 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1021 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1081 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1141 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1201 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1261 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1321 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1381 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1441 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     1501 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1561 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1621 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg
     1681 cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact
     1741 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     1801 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     1861 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     1921 ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca
     1981 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2041 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2101 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2161 tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     2221 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     2281 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     2341 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     2401 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2461 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     2521 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2581 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     2641 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2701 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     2761 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     2821 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     2881 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     2941 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3001 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3061 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3121 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3181 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3241 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3301 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3361 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3421 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3481 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3541 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3601 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3661 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3721 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3781 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3841 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3901 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3961 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4021 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4081 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4141 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4201 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4261 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4321 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4381 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4441 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4501 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4561 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4621 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4681 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4741 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4801 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4861 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4921 tcacatgt
//
