LOCUS       sequence-470639-        5324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0887m 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 5324)
  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 5324)
  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: AiP13375 -
            pAAV-AiE0887m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(226..1017,1018..1086))
                     /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     849..868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1087..1191)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1259..1277
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1315..1337)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1437..1456
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1473..1928)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1610..1631)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1822..1841
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2003..2143
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2014..2143
                     /label=AAV2 ITR
     polyA_signal    2261..2372
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2559..2579
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2666..3043)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(3074..3139)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(3152..3208)
                     /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             complement(join(3218..3928,3929..3931,3932..3934))
                     /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(3260..3281)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3609..3628
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      3931..3940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4600..4729
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4791..5324,1..55))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4871..4890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
       61 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      121 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      181 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      241 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
      301 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
      361 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
      421 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
      481 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
      541 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
      601 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
      661 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
      721 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
      781 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
      841 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
      901 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
      961 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1021 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1081 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1141 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1201 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     1261 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     1321 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1381 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     1441 atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa
     1501 attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa
     1561 aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa
     1621 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     1681 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
     1741 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     1801 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc
     1861 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     1921 gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg
     1981 agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     2041 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     2101 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg ctcggtccgc
     2161 acgtgagatc tcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
     2221 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
     2281 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
     2341 aacaacagat ggctggcaac tagaaggcac agtcgaagat ctctcgagcc acggaattgt
     2401 cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg
     2461 ccgtggcaag aactaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
     2521 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
     2581 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatgatatc
     2641 gaattcgcgg ccgcggcgcg cctcattact tagcgctggt gtacttggtg atggccttag
     2701 taccctcgga cacggcgtgc ttggccaact ccccaggcag cagcaggcgc acggccgtct
     2761 ggatctcccg ggaggtgatg gtcgagcgct tgttgtaatg cgccaggcgg gaagcctctc
     2821 ctgcgatgcg ctcgaaaatg tcgttcacaa acgaattcat gatgcccatg gccttggacg
     2881 aaatgccggt gtcagggtgg acctgcttca gaaccttgta cacatagatg gaatagctct
     2941 ccttgcggct gcgcttgcgc ttcttgccgc ctttcttctg cgccttagtc accgccttct
     3001 tggagccctt tttcggggcg ggagcagact tcgctggctc tgggcctccg gagcctccag
     3061 atctgagtcc ggacttgtca tcgtcatcct tgtaatcgat gtcatgatct ttataatctc
     3121 cgtcatggtc tttgtagtct ccactagcac caggccctgg attctcctct acatcccctg
     3181 cctgtttcaa cagtgagaag ttggtagcac cagagccctt gtacagctcg tccatgccga
     3241 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
     3301 ggtctttgct cagcttggac tggtagctca ggtagtggtt gtcgggcagc agcacggggc
     3361 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga
     3421 tgttgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat
     3481 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     3541 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     3601 cggcgcgggt cttgtagttg ccgtcgtctt tgaagaagat ggtgcgctcc tggacgtagc
     3661 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga
     3721 agcactgcac gccgtagccc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     3781 cggtggtgca gatcagcttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     3841 cggacacgct gaacttgtgg ccgttgacat cgccgtccag ctcgaccagg atgggcacca
     3901 ccccggtgaa cagctcctcg cccttggaga ccatggtggc gaccggtagc gctagcttcg
     3961 aaagatctgg atcccagaag caaatgtaag caatagatgg ctctgccctg acttttatgc
     4021 ccagcccgag ctcaacacaa ttgtctgtat tacattgcat atttaccagg tgtatatata
     4081 gttttttagg ttataaaaat aaaaaaagta ttggaggttt aattgatttt tcttctggtt
     4141 tgtttgttat atgaggtgaa tttagttttg tgtcccttct ttacctcatg gtagcatcag
     4201 aagcagctgc tgttgccttg gccacagtac agcatctact tgttgttctg gggacattaa
     4261 tgtccaaaat tggctgagct gcagggatcc agaacttctt ggaatgcaag gctacaagcc
     4321 atgactcatg ggctgaggca cctgtgagtg taacactctt gctagtctgt aatcacatgc
     4381 cctgtgtctc ttattaacca tgttctgacc ctaagggagg gcctggggac ctttagcact
     4441 actaaagtgc caagattaag gccagggcac atgaattata tgaagtcagg gctcagttct
     4501 agacttctaa aatctagaca ctcactgcct atgtgggggc tttctttgga aggaagaata
     4561 aggcattcct gaccaaatga actctacgcg tgcggccgca ggaaccccta gtgatggagt
     4621 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4681 gacgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca
     4741 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4801 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4861 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4921 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4981 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5041 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5101 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5161 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5221 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5281 agagttggta gctcttgatc cggcaaacaa accaccgctg gtag
//
