LOCUS       sequence-470759-        5261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1117m 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 5261)
  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 5261)
  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: AiP14239 -
            pAAV-AiE1117m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..45
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      114..569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(201..220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     411..432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(586..605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     705..727
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(765..783)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        851..955
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(956..1024,1025..1816)
                     /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(1174..1193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1987..2575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2476..2495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2632..2659
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2660..2687
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2694..2823
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3307..3316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3313..3315,3316..3318,3319..4029)
                     /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(3619..3638)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3966..3987
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4039..4095
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4108..4173
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4204..4581
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(4668..4688)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4875..4986
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5104..5244
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5104..5233
                     /label=AAV2 ITR
     misc_feature    join(5251..5261,1..17)
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 aattgcccat tatacgcgcg tataatggac tattgtgtgc tgatatgatg cggtattttc
       61 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
      121 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      181 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      241 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      301 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
      361 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      421 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
      481 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      541 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
      601 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      661 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
      721 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
      781 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
      841 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      901 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      961 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1021 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1081 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1141 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1201 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1261 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1321 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1381 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1441 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1501 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1561 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1621 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1681 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1741 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1801 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1861 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1921 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1981 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2041 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2101 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2221 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2281 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2341 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2401 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2461 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2521 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2581 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc atatcagcac
     2641 acaattgccc attatacgcg cgtataatgg actattgtgt gctgataaca tgtcctgcag
     2701 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
     2761 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     2821 cctgcggccg cacgcgttgc taggtaccag cctgttagtt tacctgagct caaccttaga
     2881 tcaggtcctc taaaatagag tttgacctta aaagatttgg cctccagcaa ccaatcctga
     2941 ttcattacat aaaaaaacat ccctccccag gggtgaccca agatctgatc ctgcccctcc
     3001 tcttccttcc cctcccctcc cctccagttc ctctgtcagg aggtctgaag tgttctgatc
     3061 taagatatgg atatcataaa gtcacaccat gtgacagatc tcacaataga gtttgactgg
     3121 ggaggttggc ggatccctgg ggcaggcggt gaagatgcct gactgctaac accaaacctt
     3181 tctattttgt agtacaagtc cagcaaaggt ggtgagctcg ggctgggcat aaaagtcagg
     3241 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
     3301 ccggtcgcca ccatggtctc caagggcgag gagctgttca ccggggtggt gcccatcctg
     3361 gtcgagctgg acggcgatgt caacggccac aagttcagcg tgtccggcga gggcgagggc
     3421 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     3481 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     3541 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3601 cgcaccatct tcttcaaaga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3661 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3721 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     3781 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     3841 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3901 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     3961 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4021 ctgtacaagg gctctggtgc taccaacttc tcactgttga aacaggcagg ggatgtagag
     4081 gagaatccag ggcctggtgc tagtggagac tacaaagacc atgacggaga ttataaagat
     4141 catgacatcg attacaagga tgacgatgac aagtccggac tcagatctgg aggctccgga
     4201 ggcccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact
     4261 aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag ctattccatc
     4321 tatgtgtata aggttctgaa gcaggtccac cctgacaccg gcatttcgtc caaggccatg
     4381 ggcatcatga actcgtttgt gaacgacatt ttcgagcgca tcgcaggtga ggcttcccgc
     4441 ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac ggccgtgcgc
     4501 ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa ggccatcacc
     4561 aagtacacca gcgctaagta atgaggcgcg ccgcggccgc gaattcgata tcataatcaa
     4621 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4681 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4741 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     4801 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     4861 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4921 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4981 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     5041 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     5101 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     5161 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     5221 agcgagcgcg cagctgcctg caggggcgcc tatcagcaca c
//
