LOCUS       sequence-433022-        5747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2123m 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 5747)
  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 5747)
  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: AiP1422 - pAAV-AiE2123m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   688..817
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(879..1467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(959..978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1638..2429,2430..2498))
                     /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     2261..2280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2499..2603)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2671..2689
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2727..2749)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2849..2868
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2885..3340)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3022..3043)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3234..3253
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3415..3555
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3426..3555
                     /label=AAV2 ITR
     polyA_signal    3673..3784
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3971..3991
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4084..4794,4795..4797,4798..4800))
                     /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(4126..4147)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4475..4494
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4734..4755
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4797..4806
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tttctctcat ttataagcat ttctgctgtg acctttgaac ttggcccaga gatgccatca
       61 gcagtaaact tgcaaaagcc ttcctcactc cctcagttgg cattgtgttc ccgtccactg
      121 gggggaaaaa aatataagaa aggatgacgg taatagtgca gatgagagcg atgaaggtca
      181 ctggtgacgc tatcacattt tacgtgtgtc ctttctgatc cttgcaaaat aaaaagaccc
      241 tttgggaaca tcggaacttc aggaccaatt agtggaaatg atgcttcttc tgtttgggca
      301 ttaaatacta ggcaggattc tctttaggca gctgatgctg gcagggaaag aggtaccgac
      361 ctaattactc ttccgcaaga gcaaaaaatg ccttttgaaa gcctctttgg caaatatgtc
      421 tcattaagga caaaaaccaa acacaaaccc tctaagaagc agaaattaag atgataggta
      481 acttgtggcc aggagacttt ctgtgccact gtgggaaatt aaaacaaagt ttgaggagta
      541 ccagggggtg acttgttttt gttgagtgtc tttcctgtag ccccgatgtt attatgggcc
      601 aactttattt tcttcattat tttggctcgg tgtcaggaaa tactttgctc cgccggcgat
      661 tattactctt acaacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct
      721 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg
      781 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa
      841 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      901 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      961 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1021 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1081 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1141 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1201 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1261 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1321 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1381 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1441 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1501 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1561 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1621 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1681 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1741 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1801 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1861 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1921 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1981 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2041 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2101 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2161 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2221 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2281 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2341 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2401 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2461 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2521 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2581 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2641 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     2701 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2761 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2821 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     2881 cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2941 tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc
     3001 gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3061 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3121 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3181 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3241 aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3301 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta
     3361 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca
     3421 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
     3481 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     3541 tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct gctattgtct
     3601 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     3661 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     3721 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     3781 acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     3841 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     3901 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     3961 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     4021 aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg cgccgtcgac
     4081 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     4141 gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt agctcaggta
     4201 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     4261 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc
     4321 gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt actccagctt
     4381 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     4441 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     4501 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     4561 cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg tggtcacgag
     4621 ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc
     4681 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     4741 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     4801 ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa tgtaagcaat
     4861 agatggctct gccctgactt ttatgcccag cccgagctcc gccggcgaaa gtaagcagac
     4921 agtatagaca ctggcaagca ggctcgtttc tgcatagcca atctttcagt ggcacagaaa
     4981 accatgctca tgggaacata caattttggt tttgaaaggg tctctttaac actctcctcc
     5041 tctttattca cctcaagcct tttgacactg ggcttctgca gtgaaaagtg gctctttaaa
     5101 gagagaagaa ggccacagcc agagcctgcc tgtcacttgg ataaaacata gtgaaatgat
     5161 agggctttga gcttcatttt caaggtacaa gtcaaaaaaa gaagaataag caaaaaaaag
     5221 ggaacttaac ccagaagaca gagcaggcag tctgttcttc cagtctggtt tctttggccc
     5281 atctgaatgg ggatggctga agctctgcct aatgattgct tggccatgtt tctaagggca
     5341 tccgtgtagc ctgatcatta cgttaaagaa ttggaaactt cctgagagtt aataaggcca
     5401 tttccagagc tttcaatgac gagaagtttg tgtaaatatt ctgctacatg cccatccccc
     5461 attcctatca tctgtttccc ttctcgtcat aagagtgcgt gatatgatac tccaaaactc
     5521 ctcttaggct gtggaaagga aaaacaaaaa caaaaacaga aacaaaaaca aaaacagtta
     5581 gcagtatata ctatatttcc agttcacttg atgaatcgtc ccgtatttcc cttgcgtctg
     5641 catcctcatc ttccaacagt aacgttttcc cccgagtact taacactgtc cctaaaatgg
     5701 acccatcaaa gaaatcaaac tcttgtatat tttatgcatt tgtgctt
//
