LOCUS       sequence-466245-        5221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2088m 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 5221)
  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 5221)
  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: AiP1362 - pAAV-AiE2088m-minBG-SYFP2-4X2C-WPRE3...
FEATURES             Location/Qualifiers
     source          1..5221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      204..792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     693..712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   854..983
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1866..1875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1872..1874,1875..1877,1878..2588)
                     /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(1917..1938)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2178..2197)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2525..2546
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2901..2921)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3108..3219
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3337..3477
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3337..3466
                     /label=AAV2 ITR
     rep_origin      3552..4007
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3639..3658)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3849..3870
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4024..4043)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4143..4165
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4203..4221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4289..4393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4394..4462,4463..5221,1..33)
                     /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(4612..4631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
       61 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
      121 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
      181 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      301 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      361 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      421 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      481 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      541 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      601 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      661 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      721 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      781 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      841 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
      901 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
      961 caactccatc actaggggtt cctgcggccg cacgcgttca ttttagtagg aaagtgggat
     1021 actatcttct aaagagactg tgcatcttat actcctatct accacatcat atttactgtc
     1081 tattcttagt tgggtctaaa tttctaagta attttaagta ttcatattct aggctactgg
     1141 gtcctataga ttaatataaa tatttgattg taagttatgc aataaaaatg aagagatttg
     1201 tttatatgca tctaaataag aacatgcaga atgtgttcgg aacttgaata tccttatctc
     1261 catgaaaatg ggaaaaataa aaataattta aacaaagtgt tgagtggaac atgtgtagct
     1321 gaaacacttg attttcccta aaatatctga gtaattactt gaatttccta ctttatcatt
     1381 tgtccatgga cagctggtat caggctataa aaacaacaat aattagaggc ccaaatagtt
     1441 cttttaatta ccctaaaagt ctgaaattca cgtttttgtt gtctagaagt attctaatca
     1501 tgtttaacag ctgattagca attggtccat ttttatgttg tagaaaagtg aaatgtgaat
     1561 ggaaaattga agaataattc caaaccaagg taaattatta ctgcttttcc ctgcaattgt
     1621 tagttgaaag gtgtgatttg agataagatc atttacactt aaaagtgaat agaagaaata
     1681 taataagagg gaataggcct gtgaagttgt caagaataaa ttttaaaata aatgcagcat
     1741 tttttgcatt tatgtcatag tctgtttaag cagagctcgg gctgggcata aaagtcaggg
     1801 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
     1861 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1921 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1981 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     2041 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     2101 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2161 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2221 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2281 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     2341 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     2401 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2461 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     2521 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2581 tgtacaagta agtcgacggc gcgccgcggc cttaaagaga ccggttcact gtgacagtaa
     2641 aagagaccgg ttcactgtga gaatgaaaga gaccggttca ctgtgatcgg aaaagagacc
     2701 ggttcactgt gagcggcctt gaaacccagc agacaatgta gctcagtaga aacccagcag
     2761 acaatgtagc tgaatggaaa cccagcagac aatgtagctt cggagaaacc cagcagacaa
     2821 tgtagctgtc gacgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga
     2881 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2941 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3001 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3061 gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag
     3121 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3181 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3241 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3301 caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3361 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3421 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3481 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3541 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3601 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3661 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3721 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3781 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3841 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3901 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3961 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4021 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4081 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4141 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4201 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4261 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4621 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4921 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5101 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5221 t
//
