LOCUS       sequence-425458-        4767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2004m 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 4767)
  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 4767)
  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: AiP1492 - pAAV-AiE2004m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4767
                     /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      801..810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(807..809,810..812,813..1523)
                     /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(852..873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1113..1132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1460..1481
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1588..1608)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1795..1906
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2024..2164
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2024..2153
                     /label=AAV2 ITR
     rep_origin      2245..2700
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2332..2351)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2542..2563
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2717..2736)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2836..2858
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2896..2914)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2982..3086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3087..3155,3156..3947)
                     /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(3305..3324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4118..4706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4607..4626
                     /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 gcggccgcgc atgaaacata agccttgggg gcaatcaaac tattacattg
      181 agtccttgga tttgctacaa attacatttt aaatgcaatc attttataaa agcttcaaca
      241 ctcacacttg gaagcgttac cctgttgaat atcactgact cactaacttg cattgccatg
      301 ctaacttgct ttcagagaga tctcagaaca catcatcttc tgctatttca atacatgcac
      361 attaatttcc tatcaacgtg tgctgatcag gaactctgta atctggcacc ggtgtttatt
      421 tttattcctg tctattcctg ttggctcacg aaaagattgt ttgagcaagt gttttatggt
      481 gagttgtatc atatgtacat tgatttaatc tgcccacatt cagttctaca agcggagcca
      541 aaaaaataga gacaagcata attttcattc aacatgagcc cctcaatgca agccaagtac
      601 ctcatctggt gctcagctaa agcaacagca atctgttcca ccctggagac acaactggcc
      661 acagaaaact tagtgaaaag aggcaatgct atgcacagga caaatgagct cgggctgggc
      721 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
      781 aagctagcga gctcggatcc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      841 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      901 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
      961 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct
     1021 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1081 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1141 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1201 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1261 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     1321 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1381 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     1441 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1501 tcggcatgga cgagctatac aagtaagata tcataatcaa cctctggatt acaaaatttg
     1561 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1621 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1681 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1741 gacaggggct cggctgttgg gcactgacaa ttccgtggct cgagagatct tcgactgtgc
     1801 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1861 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1921 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1981 acaatagcag gcatgagatc tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     2041 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     2101 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     2161 cagggctagc ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2221 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2281 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2341 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2401 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2461 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2521 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2581 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2641 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2701 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2761 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2821 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2881 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2941 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3001 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3061 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3121 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3181 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3241 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3301 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3361 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3421 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3481 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3541 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3601 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3661 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3721 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3961 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4021 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4081 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4141 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4201 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4261 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4321 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4381 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4441 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4501 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4561 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4621 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4681 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4741 ccttttgctg gccttttgct cacatgt
//
