LOCUS       sequence-462825-        5666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1115m 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 5666)
  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 5666)
  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: AiP14237 -
            pAAV-AiE1115m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1025..1027,1028..1030,1031..1741)
                     /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(1331..1350)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1678..1699
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1751..1807
                     /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             1820..1885
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1916..2293
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2380..2400)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2587..2698
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2816..2956
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2816..2945
                     /label=AAV2 ITR
     misc_feature    2963..2990
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2991..3018
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3087..3542
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3174..3193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3384..3405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3559..3578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3678..3700
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3738..3756)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3824..3928
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3929..3997,3998..4789)
                     /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(4147..4166)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4960..5548
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5449..5468
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5605..5632
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5633..5660
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaatcca ggacaagctt gagggtcctc tctgagctca
      181 gtagcctgct tctgaatact ctctcaggtc tccatacagt ggtttccaca tgtggcctgg
      241 acacacagca ggaaggcccc ttaaagacag cttggagggc cggatagctc cttggattcc
      301 ctgatcacca gttgcacccc agagctaatc agcccttcaa gcaactttct accagcctct
      361 accacccacc agcccctatc atccaccacc acacccttac aagccccacc cactcaccag
      421 cccccactca ccagtccccc acaggtcctg gaccacacgt gcctgaccct gaagcctgag
      481 ccaggaattt ccggccattg tccagactag agctggctca ggaagccgtg atcttcagaa
      541 aagggtgata ttgggggagg ggctttgggg acatcctggc tgagtccaga aggggccatg
      601 tgagatggca tggggtttgg gtatgcgggt gtgggggggt gtcttcctcc ttgtgcacca
      661 tgacttaaga aggcagggca ttccacttct gcctggtaag acccagggcc atcttcatat
      721 ggtcactctc aaggcaaggg aagagaggaa gagcggggat aggacactgg ataagcccag
      781 gaagccatgc tggcgacaca gaagagacct gggctcagag ttgctggctt accctagtat
      841 ttgaatccta gaactggagt caggagctca gagaagtcag aggacagcac tagccacaca
      901 cagggacttg tgttctaatc tggctgagct cgggctgggc ataaaagtca gggcagagcc
      961 atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc
     1021 caccatggtc tccaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1081 ggacggcgat gtcaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1141 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
     1201 caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat
     1261 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1321 cttcttcaaa gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1381 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1441 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     1501 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     1561 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1621 ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1681 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1741 gggctctggt gctaccaact tctcactgtt gaaacaggca ggggatgtag aggagaatcc
     1801 agggcctggt gctagtggag actacaaaga ccatgacgga gattataaag atcatgacat
     1861 cgattacaag gatgacgatg acaagtccgg actcagatct ggaggctccg gaggcccaga
     1921 gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca
     1981 gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta
     2041 taaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat
     2101 gaactcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca
     2161 ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct
     2221 gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac
     2281 cagcgctaag taatgaggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga
     2341 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2401 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2461 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     2521 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat
     2581 cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2641 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2701 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2761 aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa
     2821 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2881 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     2941 cgcagctgcc tgcaggggcg cctatcagca cacaattgcc cattatacgc gcgtataatg
     3001 gactattgtg tgctgatatg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3061 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3121 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     3181 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3241 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3301 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3361 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3421 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3481 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3541 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3601 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3661 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3721 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3781 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3841 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3901 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3961 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4021 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4081 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4141 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4201 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4261 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4321 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4381 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4441 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4501 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4561 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4621 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4681 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4741 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4801 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4861 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4921 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4981 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5041 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5101 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5161 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5221 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5281 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5341 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5401 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5461 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5521 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5581 ggccttttgc tggccttttg ctcatatcag cacacaattg cccattatac gcgcgtataa
     5641 tggactattg tgtgctgata acatgt
//
