LOCUS       sequence-465059-        5078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2434m 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 5078)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5078)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP1758 - pAAV-AiE2434m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5078
                     /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      1090..1099
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1096..1098,1099..1101,1102..1812)
                     /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(1141..1162)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1402..1421)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1749..1770
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1905..1925)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2112..2223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2341..2481
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2341..2470
                     /label=AAV2 ITR
     rep_origin      2556..3011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2643..2662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2853..2874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3028..3047)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3147..3169
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3207..3225)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3293..3397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3398..3466,3467..4258)
                     /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(3616..3635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4429..5017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4918..4937
                     /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 gcgttgtaag agtaataatc gcaaattttc actttcactc
      181 tctatagcag acagtgaagc acttcccaga tgagctatgg tgtctggggc tggaaggaag
      241 gtggtaccct ctaagaaggt aatatccaca agatgaatcc atgtgtcagg agaggcagat
      301 ctggaattca accaggttgg aggagctaat ggcttatgcc ttttagttac ctacctcctt
      361 tgatgggttt ctgataccta ctgtctagaa aggcacctaa gggggctttc tcgcatccta
      421 gcgaccagat cttttccaac actaaatcac tgcgtctctg atgtcagtta agtatttggc
      481 agcttgattt aattctaaca ctacatccca ttagcccaga ctcccacagg tagaaggctt
      541 agtcctaaaa taggacctca cttcagatgc cagtcacaag tccaacctga gctccagaaa
      601 acccacctgt aaatttcagg gttccattaa gagcttctcg ggttcagaaa ttccctagag
      661 tactttgcag aacttgagaa atcactttat ttaccagcat gcagtgaata atacaattca
      721 ggaacggcca gatggaaaca acacaggggg tgaagtcagc agagaaagtg ggcccagagt
      781 gctgagcctt gtctgaacat gtccccctcc cacagctcca cattcaccaa ccggaagctc
      841 ctccagcctc aacccttagg aattttaatg gaggtttcat cttgttggca caatcgatgc
      901 agttattgtc cagcagtgat tgaaatcagt caccagccct tccgctcgac tcgagactcg
      961 agccccgggc taccttgcca gcagagtctt gccccggagc tcgggctggg cataaaagtc
     1021 agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg
     1081 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1141 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1201 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1261 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     1321 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1381 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1441 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1501 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1561 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1621 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1681 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     1741 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1801 gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct
     1861 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1921 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1981 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     2041 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga
     2101 gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2161 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2221 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2281 gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc
     2341 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2401 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     2461 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     2521 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     2581 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     2641 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2701 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2761 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2821 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2881 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     2941 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3001 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3061 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3121 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3181 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3241 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3301 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3361 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3421 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3481 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3541 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3601 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3661 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3721 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3781 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3841 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3901 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3961 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4021 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4081 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4141 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4201 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4261 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4321 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4381 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4441 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4501 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4561 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4621 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4681 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4741 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4801 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4861 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4921 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4981 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5041 acggttcctg gccttttgct ggccttttgc tcacatgt
//
