LOCUS       sequence-470643-        5347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0890m 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 5347)
  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 5347)
  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: AiP13407 -
            pAAV-AiE0890m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      150..159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(156..158,159..161,162..872)
                     /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(462..481)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     809..830
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             882..938
                     /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             951..1016
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1047..1424
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1511..1531)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1718..1829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1947..2087
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1947..2076
                     /label=AAV2 ITR
     rep_origin      2162..2617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2249..2268)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2459..2480
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2634..2653)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2753..2775
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2813..2831)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2899..3003
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3004..3072,3073..3864)
                     /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(3222..3241)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4035..4623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4524..4543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4685..4814
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gctcagtctc tttaatcata cctagtcagt atctgtcacg gggataaact aaggcagagc
       61 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
      121 cagatctttc gaagctagcg ctaccggtcg ccaccatggt ctccaagggc gaggagctgt
      181 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga tgtcaacggc cacaagttca
      241 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
      301 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
      361 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      421 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa agacgacggc aactacaaga
      481 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      541 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
      601 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
      661 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
      721 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
      781 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
      841 ggatcactct cggcatggac gagctgtaca agggctctgg tgctaccaac ttctcactgt
      901 tgaaacaggc aggggatgta gaggagaatc cagggcctgg tgctagtgga gactacaaag
      961 accatgacgg agattataaa gatcatgaca tcgattacaa ggatgacgat gacaagtccg
     1021 gactcagatc tggaggctcc ggaggcccag agccagcgaa gtctgctccc gccccgaaaa
     1081 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca
     1141 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca
     1201 ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc
     1261 gcatcgcagg agaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctccc
     1321 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt
     1381 ccgagggtac taaggccatc accaagtaca ccagcgctaa gtaatgaggc gcgccgcggc
     1441 cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1501 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1561 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     1621 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     1681 tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca
     1741 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1801 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1861 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag
     1921 atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     1981 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     2041 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     2101 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     2161 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2221 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2281 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2341 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     2401 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2461 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     2521 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     2581 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     2641 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2701 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2761 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2821 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2881 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2941 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3001 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3061 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3121 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3181 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3241 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3301 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3361 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3421 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3481 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3541 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3601 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3661 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3721 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3781 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3841 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3901 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3961 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4021 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4081 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4141 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4201 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4261 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4321 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4381 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4441 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4501 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4561 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4621 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4681 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
     4741 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     4801 cactaggggt tcctgcggcc gcacgcgttg agcttcaacc aaatcaggca ttgatggatt
     4861 ttatagtttg attaacaaag ataatagcaa accccagatt tagtttaaac ataaaaagta
     4921 ttaaggttgt atcctgcttg tatagcatat gcaaatgacc tcgtttctgc tactgcattt
     4981 ggaaatgtag cagaagaaaa aaaaaaggca cttcaattgc agctctcatc agttattcac
     5041 tgtatccagg cctctcaatt gtgttctttt ctttaatgca atagcaagca gcaatcaccc
     5101 agctgtgctt ggtagagtga actatataca catctatatt gagatttcat acacacataa
     5161 cataaaagcg agagaaaaag cctcaagaat gtttggccca ttgcaaatca cacaaaagga
     5221 ctaatgaatc tctctccaaa tggatctgta gtgaccatct gtaagccttg attgattcat
     5281 attccataac ggtatcagca tccaggaagt gattacttca aggtgcaaca caacttcccc
     5341 tatgaaa
//
