LOCUS       sequence-441078-        4788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2333m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4788)
  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 4788)
  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: AiP1653 - pAAV-AiE2333m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      800..809
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(806..808,809..811,812..1522)
                     /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(851..872)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1112..1131)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1459..1480
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1615..1635)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1822..1933
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2051..2191
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2051..2180
                     /label=AAV2 ITR
     rep_origin      2266..2721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2353..2372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2563..2584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2738..2757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2857..2879
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2917..2935)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3003..3107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3108..3176,3177..3968)
                     /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(3326..3345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4139..4727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4628..4647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      121 cggccgcacg cgttgtaaga gtaataatcg agtcttctca aagataatgt tctttaggtc
      181 acaatgtaag aatatatggt agtgggagaa acttttggtg ggataggagg ctgaaaaatg
      241 agaagctaca taagtactct agttcttcag tctttctgta actgtttttt tagttgacat
      301 tttttttttt ttttactgca aagaaggata tatttatata gctgtcaaag aggagactcg
      361 ccacatatgc catctgccct tcccagaacc cacacactga gctgtgtagg tggaacctct
      421 tctggctgac tttctaaggt tcacacatgt catctttcac atcaccattt atgagaaatt
      481 tcatatcagc agaattttat gtgttgttta gaaaacacac tccttgacat tatctatttc
      541 ctcagattag gattcctatt ttgatttatt ttacaatctg aaagttaccc caaactgaag
      601 aggaggaagg agaataagga gaaggaagaa gagaaggaag aggagaagga ggagtaggag
      661 aggaggtgaa gtaggaggaa gaggaagagg aagattgtga tgaacggagc tcgggctggg
      721 cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc
      781 gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      841 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      901 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
      961 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt
     1021 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1081 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1141 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1201 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1261 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     1321 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1381 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     1441 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1501 cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca
     1561 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1621 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1681 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     1741 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     1801 cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1861 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1921 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1981 aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc
     2041 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2101 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2161 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     2221 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     2281 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2341 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2401 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2461 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2521 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2581 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     2641 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2701 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     2761 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2821 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2881 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     2941 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3001 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3061 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3121 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3181 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3241 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3301 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3361 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3421 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3481 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3541 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3601 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3661 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3721 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3781 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3841 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3901 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3961 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4021 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4081 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4141 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4201 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4261 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4321 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4381 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4441 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4501 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4561 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4621 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4681 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4741 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
