LOCUS       sequence-449742-        5468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2118m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in forebrain astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5468)
  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 5468)
  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: AiP14480 -
            pAAV-AiE2118m_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..5468
                     /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      1480..1489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1486..1488,1489..1491,1492..2202)
                     /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(1531..1552)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1792..1811)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2139..2160
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2295..2315)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2502..2613
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2731..2871
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2731..2860
                     /label=AAV2 ITR
     rep_origin      2946..3401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3033..3052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3243..3264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3418..3437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3537..3559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3597..3615)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3683..3787
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3788..3856,3857..4648)
                     /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(4006..4025)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4819..5407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5308..5327
                     /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 gcgttgtgaa tctggatgcg tattgaaaga aacattagtc
      181 cttttgtcag cttggcaagt ctattgttct gagccaggcc caaccaatta acatcttttg
      241 ccaatccctg tcagcagggg cttgcaagga ggagagaaag gggggccggt cagatgctaa
      301 tttagataca attgtgtcag ggtgcatggg aggggtaact ctgaagaaga gactccagtt
      361 taatgaagca aacagcttgg gtcccagtga ccctgcaggg gtaattttaa aaggcccctc
      421 tgtgttcctg tgccaaattg cagagggcca gcaacaagaa gcccttcaaa aaaaaaaaaa
      481 gagggagcag gataatcatc cacttaatct gctcattaaa caaatctctc aggcggactt
      541 aacgtgaatt agcacttctg tgaatctgga tgcgtattga aagaaacatt agtccttttg
      601 tcagcttggc aagtctattg ttctgagcca ggcccaacca attaacatct tttgccaatc
      661 cctgtcagca ggggcttgca aggaggagag aaaggggggc cggtcagatg ctaatttaga
      721 tacaattgtg tcagggtgca tgggaggggt aactctgaag aagagactcc agtttaatga
      781 agcaaacagc ttgggtccca gtgaccctgc aggggtaatt ttaaaaggcc cctctgtgtt
      841 cctgtgccaa attgcagagg gccagcaaca agaagccctt caaaaaaaaa aaaagaggga
      901 gcaggataat catccactta atctgctcat taaacaaatc tctcaggcgg acttaacgtg
      961 aattagcact tctgtgaatc tggatgcgta ttgaaagaaa cattagtcct tttgtcagct
     1021 tggcaagtct attgttctga gccaggccca accaattaac atcttttgcc aatccctgtc
     1081 agcaggggct tgcaaggagg agagaaaggg gggccggtca gatgctaatt tagatacaat
     1141 tgtgtcaggg tgcatgggag gggtaactct gaagaagaga ctccagttta atgaagcaaa
     1201 cagcttgggt cccagtgacc ctgcaggggt aattttaaaa ggcccctctg tgttcctgtg
     1261 ccaaattgca gagggccagc aacaagaagc ccttcaaaaa aaaaaaaaga gggagcagga
     1321 taatcatcca cttaatctgc tcattaaaca aatctctcag gcggacttaa cgtgaattag
     1381 cacttcgagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
     1441 ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc
     1501 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1561 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1621 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1681 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1741 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1801 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1861 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1921 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1981 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     2041 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     2101 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2161 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg
     2221 cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact
     2281 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2341 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     2401 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2461 ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca
     2521 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2581 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2641 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2701 tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     2761 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     2821 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     2881 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     2941 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3001 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     3061 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3121 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     3181 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3241 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     3301 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     3361 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     3421 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3481 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3541 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3601 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3661 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3721 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3781 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3841 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3901 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3961 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4021 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4081 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4141 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4201 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4261 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4321 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4381 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4441 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4501 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4561 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4621 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4681 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4741 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4801 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4861 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4921 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4981 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5041 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5101 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5161 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5221 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5281 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5341 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5401 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5461 tcacatgt
//
