LOCUS       sequence-461889-        5563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1125m 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 5563)
  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 5563)
  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: AiP14259 -
            pAAV-AiE1125m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5563
                     /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      916..925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(922..924,925..927,928..1638)
                     /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(1228..1247)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1575..1596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1648..1704
                     /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             1717..1782
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1813..2190
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2277..2297)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2484..2595
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2713..2853
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2713..2842
                     /label=AAV2 ITR
     misc_feature    2860..2887
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2888..2915
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2984..3439
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3071..3090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3281..3302
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3456..3475)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3575..3597
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3635..3653)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3721..3825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3826..3894,3895..4686)
                     /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(4044..4063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4857..5445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5346..5365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5502..5529
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5530..5557
                     /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 gcgtagaacc tgggagcaga ttaatggcag ctgctggtct
      181 agtttctgaa aaggactcca atgtcccatg tcctacttgg cctcaaggac ttaggtatca
      241 agtcacagcc ctaaagtccc aaacctcctg acctagtgcg aggggaaatg ctagaatgtg
      301 ctagaagccc cgtgactcac tggattcctt ctgcagttcg ttccacaccc agatccgtcc
      361 tttgcaggtg accagggact accgcttccg agccggcacc aatggggacc gagaaagggc
      421 cccagcaaca gagagggatt ggctgaggcc gcctacgcga gatgggctgt tctgggctta
      481 ggcccccagc tgcgtgatgg gtggggggag ggggggggag atgggtggct gccgcctgca
      541 tgtccctgca gctgggtcat cccggtctga aggtcacacc tggtgtgtga ctggactcgg
      601 taataggggg cgggacaaag cacaaaggtc agttggtggt gtttttctcc tcataaggaa
      661 gtgtggagcc agaagtctgt tttgccacgg gttagctcat cagatgaggt gaccaaggcc
      721 aaaggcaggc ctcagcttgt agagcaaaga gagggagaaa aacagggcgc agtgttctgc
      781 ttcctgttta tgaacaagca tagagaagcc ggacttggct cagagctcgg gctgggcata
      841 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
      901 ctagcgctac cggtcgccac catggtctcc aagggcgagg agctgttcac cggggtggtg
      961 cccatcctgg tcgagctgga cggcgatgtc aacggccaca agttcagcgt gtccggcgag
     1021 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     1081 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     1141 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1201 gtccaggagc gcaccatctt cttcaaagac gacggcaact acaagacccg cgccgaggtg
     1261 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1321 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1381 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1441 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1501 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     1561 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1621 atggacgagc tgtacaaggg ctctggtgct accaacttct cactgttgaa acaggcaggg
     1681 gatgtagagg agaatccagg gcctggtgct agtggagact acaaagacca tgacggagat
     1741 tataaagatc atgacatcga ttacaaggat gacgatgaca agtccggact cagatctgga
     1801 ggctccggag gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag
     1861 gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc
     1921 tattccatct atgtgtataa ggttctgaag caggtccacc ctgacaccgg catttcgtcc
     1981 aaggccatgg gcatcatgaa ctcgtttgtg aacgacattt tcgagcgcat cgcaggtgag
     2041 gcttcccgcc tggcgcatta caacaagcgc tcgaccatca cctccaggga gatccagacg
     2101 gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag
     2161 gccatcacca agtacaccag cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat
     2221 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2281 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2341 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     2401 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2461 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2521 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2581 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2641 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     2701 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2761 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2821 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct atcagcacac aattgcccat
     2881 tatacgcgcg tataatggac tattgtgtgc tgatatgatg cggtattttc tccttacgca
     2941 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     3001 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3061 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3121 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3181 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3241 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3301 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     3361 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3421 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3481 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3541 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3601 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     3661 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     3721 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3781 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3841 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3901 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3961 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4021 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4081 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4141 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4201 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4261 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4321 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4381 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4441 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4501 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4561 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4621 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4681 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4741 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4801 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4861 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4921 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4981 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5041 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5101 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5161 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5221 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5281 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5341 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5401 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5461 gcctttttac ggttcctggc cttttgctgg ccttttgctc atatcagcac acaattgccc
     5521 attatacgcg cgtataatgg actattgtgt gctgataaca tgt
//
