LOCUS       sequence-460613-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0128h_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 5510)
  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 5510)
  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: AiP12570 -
            pAAV-AiE0128h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5510
                     /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      1522..1531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1528..1530,1531..1533,1534..2244)
                     /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(1573..1594)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1834..1853)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2181..2202
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2337..2357)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2544..2655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2773..2913
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2773..2902
                     /label=AAV2 ITR
     rep_origin      2988..3443
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3075..3094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3285..3306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3460..3479)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3579..3601
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3639..3657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3725..3829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3830..3898,3899..4690)
                     /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(4048..4067)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4861..5449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5350..5369
                     /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 gcgtctagta tcatgaggag gatctcaaag cagggaaggg
      181 aagccagctg agggacccct gagaggagat caggggataa atactttgca acagaatccc
      241 catgggaaac atgacaaaat tagtaacagg aatggagcct ccctcagagg atacatgaat
      301 cgttgagttg aaaataacca caaggtcatc aaatccagct gccaccaact gcctgaatat
      361 tttcaacaaa tttttgaaca agcagtcctc cagcctcctc cagctgcatc cagcagctca
      421 gtgctccacc tgacagcctc ttctgatagc gtgaaggctc agaaagcccc tgtggggccc
      481 agcacatcct ggggcaggcc tctagtgggc agagggcagc atactggggt ctggatctaa
      541 cacactcctc catccaggtg gtatatttat agctagtatc atgaggagga tctcaaagca
      601 gggaagggaa gccagctgag ggacccctga gaggagatca ggggataaat actttgcaac
      661 agaatcccca tgggaaacat gacaaaatta gtaacaggaa tggagcctcc ctcagaggat
      721 acatgaatcg ttgagttgaa aataaccaca aggtcatcaa atccagctgc caccaactgc
      781 ctgaatattt tcaacaaatt tttgaacaag cagtcctcca gcctcctcca gctgcatcca
      841 gcagctcagt gctccacctg acagcctctt ctgatagcgt gaaggctcag aaagcccctg
      901 tggggcccag cacatcctgg ggcaggcctc tagtgggcag agggcagcat actggggtct
      961 ggatctaaca cactcctcca tccaggtggt atatttatag ctagtatcat gaggaggatc
     1021 tcaaagcagg gaagggaagc cagctgaggg acccctgaga ggagatcagg ggataaatac
     1081 tttgcaacag aatccccatg ggaaacatga caaaattagt aacaggaatg gagcctccct
     1141 cagaggatac atgaatcgtt gagttgaaaa taaccacaag gtcatcaaat ccagctgcca
     1201 ccaactgcct gaatattttc aacaaatttt tgaacaagca gtcctccagc ctcctccagc
     1261 tgcatccagc agctcagtgc tccacctgac agcctcttct gatagcgtga aggctcagaa
     1321 agcccctgtg gggcccagca catcctgggg caggcctcta gtgggcagag ggcagcatac
     1381 tggggtctgg atctaacaca ctcctccatc caggtggtat atttatagga gctcgggctg
     1441 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt
     1501 tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1561 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1621 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1681 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     1741 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1801 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1861 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1921 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1981 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     2041 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2101 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     2161 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2221 ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg aattcgatat
     2281 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2341 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2401 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     2461 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2521 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2581 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2641 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2701 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     2761 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2821 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2881 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2941 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3001 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3061 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3121 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3181 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3241 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3301 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3361 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3421 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3481 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3541 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3601 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3661 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3721 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3781 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3841 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3901 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3961 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4021 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4081 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4141 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4201 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4261 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4321 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4381 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4441 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4501 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4561 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4621 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4681 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4741 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4801 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4981 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
