LOCUS       sequence-432587-        4641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2550m 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 4641)
  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 4641)
  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: AiP1956 - pAAV-AiE2550m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..48)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(708..1499,1500..1568))
                     /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     1331..1350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1569..1673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1741..1759
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1797..1819)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1919..1938
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1955..2410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2092..2113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2304..2323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2485..2625
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2496..2625
                     /label=AAV2 ITR
     polyA_signal    2743..2854
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3041..3061
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3154..3864,3865..3867,3868..3870))
                     /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(3196..3217)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3545..3564
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3804..3825
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3867..3876
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4399..4528
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4590..4641,1..537))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
       61 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      121 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      181 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      241 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      301 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      361 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      421 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      481 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      541 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      601 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      661 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      721 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      781 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      841 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      901 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      961 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1021 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1081 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1141 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1201 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1261 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1321 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1381 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1441 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1501 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1561 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1621 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1681 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     1741 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     1801 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     1861 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     1921 gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta
     1981 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc
     2041 aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg
     2101 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2161 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
     2221 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2281 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg
     2341 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2401 cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa
     2461 ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg
     2521 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     2581 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc
     2641 gcacgtgaga tctcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     2701 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     2761 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     2821 caaacaacag atggctggca actagaaggc acagtcgaag atctctcgag ccacggaatt
     2881 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     2941 cgccgtggca agaactaacc aggatttata caaggaggag aaaatgaaag ccatacggga
     3001 agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac
     3061 atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt gattatgata
     3121 tcgaattcgc ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca tgccgagagt
     3181 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     3241 tttgctcagc ttggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc
     3301 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt
     3361 gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac
     3421 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     3481 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     3541 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     3601 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca
     3661 ctgcacgccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     3721 ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     3781 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     3841 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag
     3901 atctggatcc cagaagcaaa tgtaagcaat agatggctct gccctgactt ttatgcccag
     3961 cccgagctcc gagagacaaa atgatcactg gatccagggt atcccagact acatagctag
     4021 acaccaatcc aatttaaagc acaaaatcaa aaactagagg aaaatagtcc ctgtcagtgc
     4081 ctttcctgtt cctatgctgc atataaactg ctgtgagtca tgtcagactt gagctcatta
     4141 tgtagtcact ctggatttgt tgcttataca gtgaaccaac tccaagaaat agaaacactg
     4201 cctactctgt taaaaatatg aaaaattcca gtcttccatc aagcatggga atccatttat
     4261 ttacctagca atattttagg ctagtgtgca gagtaactag tttctcagga tatcttcata
     4321 tgtaggcgtc acactttttc tctttttctc ttctttccag ttactgtcga ttattactct
     4381 tacaacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4441 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg
     4501 agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca aaaggccagg
     4561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4621 cacaaaaatc gacgctcaag t
//
