LOCUS       sequence-466351-        4696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2410m 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 4696)
  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 4696)
  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: AiP20021 - pAAV-AiE2410m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      534..1122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1023..1042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1184..1313
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1708..1725
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     regulatory      1891..1900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1897..1899,1900..1902,1903..2613)
                     /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(1942..1963)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2203..2222)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2550..2571
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2706..2726)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2913..3024
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3142..3282
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3142..3271
                     /label=AAV2 ITR
     rep_origin      3357..3812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3444..3463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3654..3675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3829..3848)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3948..3970
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4008..4026)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4094..4198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4199..4267,4268..4696,1..363)
                     /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(4417..4436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
       61 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
      121 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
      181 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
      241 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      301 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
      361 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
      421 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      481 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      541 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      601 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      661 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
      721 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      781 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      841 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      901 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      961 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1021 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1081 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1141 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     1201 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     1261 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     1321 cacgcgtggt gttggccctg tgagcacact tccttcagca cttccaatca tttgtttgga
     1381 acacatctga gttcctaccc tcaggagcaa gggctcaaat tctaggaagg ctcacaaatc
     1441 cagccttgcc ctaggctcag gaagcacaga tcacatccct ggcatgggac tcttcctgct
     1501 tcgagctcag ggcttagtcc aacaccacaa ggcagatttt gcagccagaa acagatataa
     1561 atttggctga gcagagccag tggaagcaaa atagattctg taatgtacag aatatgcagc
     1621 agatggctct gcttcctacc agccctgtaa gactgattaa cacagcccac tgccctccat
     1681 caccaccacc atccttatta ctaccagcat cagcaccagc accagcacat cgtattaaaa
     1741 gtcttctggt acatacatgg cccttgcaaa atgctccatg cctgacatcc gccggcggag
     1801 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
     1861 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
     1921 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1981 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
     2041 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
     2101 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2161 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2221 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2281 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2341 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     2401 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     2461 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     2521 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2581 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     2641 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2701 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2761 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2821 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2881 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2941 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3001 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3061 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     3121 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     3181 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3241 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     3301 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     3361 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3421 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3481 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3541 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     3601 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3661 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     3721 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3781 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3841 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3901 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3961 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     4021 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4081 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4141 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4201 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4261 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4321 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4381 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4441 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4501 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4561 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4621 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4681 tcgttgggaa ccggag
//
