LOCUS       sequence-470724-        5110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0549h 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 5110)
  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 5110)
  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: AiP13652 -
            pAAV-AiE0549h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      702..1290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1191..1210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1352..1481
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1927..1936
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1933..1935,1936..1938,1939..2649)
                     /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(2239..2258)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2586..2607
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2659..2715
                     /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             2728..2793
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2824..3201
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(3288..3308)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3495..3606
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3724..3864
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3724..3853
                     /label=AAV2 ITR
     rep_origin      3939..4394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4026..4045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4236..4257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4411..4430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4530..4552
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4590..4608)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4676..4780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4781..4849,4850..5110,1..531)
                     /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(4999..5018)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
       61 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
      121 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
      181 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
      241 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
      301 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
      361 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
      421 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
      481 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
      541 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
      601 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
      661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      841 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1081 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1141 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1201 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1261 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1321 ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc
     1381 actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     1441 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca cgcgtcagaa
     1501 ggtaatatat actctgatag agttaaagta tcatgaaaga aaaagagaag aaatgcattt
     1561 ggtttgagga tctgtgaaag cttcaagcag aaggtggaat ttggaagtgg gtcttgaaaa
     1621 atgtgtggat tcctctggga agtgaattga accagtagaa ttccacactg aagaaacagc
     1681 aggcaaaggc aaggaggtgt gaaactcccc agtcttcaga agctacagtg ggccctctaa
     1741 gtctgaaaag aatgtgggtg aaaggacagt gtgagagata aggccacaaa ggaaagttag
     1801 gaccagacac cccgcattct caatgccttc tacgagctcg ggctgggcat aaaagtcagg
     1861 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
     1921 ccggtcgcca ccatggtctc caagggcgag gagctgttca ccggggtggt gcccatcctg
     1981 gtcgagctgg acggcgatgt caacggccac aagttcagcg tgtccggcga gggcgagggc
     2041 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     2101 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     2161 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2221 cgcaccatct tcttcaaaga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2281 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2341 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     2401 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     2461 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2521 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     2581 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2641 ctgtacaagg gctctggtgc taccaacttc tcactgttga aacaggcagg ggatgtagag
     2701 gagaatccag ggcctggtgc tagtggagac tacaaagacc atgacggaga ttataaagat
     2761 catgacatcg attacaagga tgacgatgac aagtccggac tcagatctgg aggctccgga
     2821 ggcccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact
     2881 aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag ctattccatc
     2941 tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc caaggccatg
     3001 ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggaga ggcttcccgc
     3061 ctggcgcatt acaacaagcg ctcgaccatc acctcccggg agatccagac ggccgtgcgc
     3121 ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa ggccatcacc
     3181 aagtacacca gcgctaagta atgaggcgcg ccgcggccgc gaattcgata tcataatcaa
     3241 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3301 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3361 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     3421 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     3481 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3541 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3601 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3661 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     3721 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3781 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3841 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     3901 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3961 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4021 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4081 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4141 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4201 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4261 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     4321 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4381 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4441 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4501 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4561 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4621 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4681 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4741 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4801 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4861 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4921 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4981 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5041 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5101 agaaaagcat
//
