LOCUS       sequence-466297-        4968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2398m 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 4968)
  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 4968)
  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: AiP1728 - pAAV-AiE2398m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   218..347
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(409..997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(489..508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1168..1959,1960..2028))
                     /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     1791..1810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2029..2133)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2201..2219
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2257..2279)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2379..2398
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2415..2870)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2552..2573)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2764..2783
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2945..3085
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2956..3085
                     /label=AAV2 ITR
     polyA_signal    3203..3314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3501..3521
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3614..4324,4325..4327,4328..4330))
                     /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(3656..3677)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4005..4024
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4264..4285
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4327..4336
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tatgcacagc actcaatgaa caaaattttg agatgaattt tcaagagtag agagaaacca
       61 atgcacccac cctgccagcg tgtttcacta tgtttccttt cttttttata cggttcagag
      121 actactctag ggaatggtgt cacccactgt gggcatagac agtgaacagt aagacaatcc
      181 attatacgat tattactctt acaacgcgtg cggccgcagg aacccctagt gatggagttg
      241 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
      301 cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa
      361 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      421 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      481 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      541 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      601 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      661 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      721 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      781 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      841 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      901 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      961 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1021 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1081 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1141 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1201 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1261 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     1321 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1381 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     1441 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     1501 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     1561 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     1621 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1681 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1741 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1801 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1861 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1921 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1981 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     2041 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2101 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     2161 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     2221 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2281 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2341 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     2401 gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     2461 ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa
     2521 agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2581 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2641 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2701 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2761 ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct
     2821 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg
     2881 ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     2941 cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
     3001 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     3061 gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct
     3121 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
     3181 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
     3241 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
     3301 actagaaggc acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag
     3361 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc
     3421 aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag
     3481 gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca
     3541 atctttcaca aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg
     3601 cgccgtcgac ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     3661 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt
     3721 agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     3781 ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg
     3841 ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt
     3901 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3961 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     4021 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     4081 agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg
     4141 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg
     4201 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     4261 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     4321 tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa
     4381 tgtaagcaat agatggctct gccctgactt ttatgcccag cccgagctcc gtatgagggt
     4441 ggcatttggg aagattcctc tgtaatgtct tggttgtccc ctccttattt gaattaattc
     4501 ctaggcctgc ctggccttta tggagatttt ggaactttaa ttttctataa aacatgggtc
     4561 aattatcagt ttattttgaa aaattttatt tgtcattaga caggaaaaga atattgtcaa
     4621 gttgtactct ttgagcccct accctggaca tcagagattc tcctttatag cctcagattt
     4681 tagttttctt cccatctctt tttcagcttt agattttttt ttcttttctc acagactgga
     4741 ggggaataac agctacagga aggagcctgg gatgtctgtt cacaccacat ccacagccaa
     4801 gagtagagag aagagtgaga gaacgctcac tcagcagtta agagcactgg ctttattcat
     4861 cagcaccatc atcacagtta acaattgtaa ctccattccc agggggttag atgccctctt
     4921 ctggcctcct caggcaccag gctcgcacat ggcacaaaga tatacaca
//
