LOCUS       sequence-470760-        5734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1118m 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 5734)
  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 5734)
  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: AiP14240 -
            pAAV-AiE1118m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      53..641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     542..561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    698..725
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    726..753
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   760..889
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1846..1855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1852..1854,1855..1857,1858..2568)
                     /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(2158..2177)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2505..2526
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2578..2634
                     /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             2647..2712
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2743..3120
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(3207..3227)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3414..3525
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3643..3783
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3643..3772
                     /label=AAV2 ITR
     misc_feature    3790..3817
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3818..3845
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3914..4369
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4001..4020)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4211..4232
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4386..4405)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4505..4527
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4565..4583)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4651..4755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4756..4824,4825..5616)
                     /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(4974..4993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
       61 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      121 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      181 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      241 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      301 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      361 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      421 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      481 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      541 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      601 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      661 ttttacggtt cctggccttt tgctggcctt ttgctcatat cagcacacaa ttgcccatta
      721 tacgcgcgta taatggacta ttgtgtgctg ataacatgtc ctgcaggcag ctgcgcgctc
      781 gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
      841 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg
      901 cgtccttcct gggcaactta agaagaatct gtctcaagat taaaaaatac aaaaaagatt
      961 gtacaagctc agtgtttagc atgcgtaagg ctctgggtcc aattcttaaa actgttataa
     1021 aatttagaaa gtttttttaa gagggaaaga aatgaaaaga agcatattag aaccagtaca
     1081 tagagatggg agcacccagt gtgacatcca agatgggagc acccagtgtg ccgtccaacc
     1141 aacacgacag catagacaca gaaatactga gggagaggaa aggcaatcag gacaggtttt
     1201 ctaaaggtct tggaggcagg tttgggggtg atatcccact gcggaactga ctctccttgt
     1261 catttctaag gtctcctcag gggctcctct tctgaagcca tccccctcct gctctggctg
     1321 atctcattgt cacccccggg ctctgttgtt gggtgtggct gtccactctt tagtcactgc
     1381 cctggctctc caggacaccc cggcaggact aggcatggac tgataagagc tgtgcttgtt
     1441 agcacagctg ttctttgcaa cacccaagcc ttgttttcaa acctgcggaa acttgggtgc
     1501 ggaagcgggg ctgcatccac ccgacttcag agttgggctc gcttccagat aaaaggaaat
     1561 tcagtcagtt ttcccagaca cgttgacaaa aatagtgttg acaggaacct catagggtct
     1621 gctggggttc cagtgtgaca tctacaaaag tcagctgggc tttgtgactg gatgctgtta
     1681 ggagtcctgt tcaaatgaaa aacttctctg gaagtggttc ttgtggaggt tctttgggga
     1741 caaggttcga cagagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
     1801 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtctcc
     1861 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgatgtc
     1921 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1981 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     2041 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     2101 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaagac
     2161 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2221 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2281 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     2341 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     2401 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2461 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2521 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg ctctggtgct
     2581 accaacttct cactgttgaa acaggcaggg gatgtagagg agaatccagg gcctggtgct
     2641 agtggagact acaaagacca tgacggagat tataaagatc atgacatcga ttacaaggat
     2701 gacgatgaca agtccggact cagatctgga ggctccggag gcccagagcc agcgaagtct
     2761 gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa gaaaggcggc
     2821 aagaagcgca agcgcagccg caaggagagc tattccatct atgtgtataa ggttctgaag
     2881 caggtccacc ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa ctcgtttgtg
     2941 aacgacattt tcgagcgcat cgcaggtgag gcttcccgcc tggcgcatta caacaagcgc
     3001 tcgaccatca cctccaggga gatccagacg gccgtgcgcc tgctgctgcc tggggagttg
     3061 gccaagcacg ccgtgtccga gggtactaag gccatcacca agtacaccag cgctaagtaa
     3121 tgaggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta caaaatttgt
     3181 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3241 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3301 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3361 acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt cgactgtgcc
     3421 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3481 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3541 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3601 caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     3661 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3721 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     3781 aggggcgcct atcagcacac aattgcccat tatacgcgcg tataatggac tattgtgtgc
     3841 tgatatgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     3901 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3961 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     4021 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4081 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4141 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4201 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4261 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4321 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4381 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4441 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4501 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4561 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4621 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4681 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4741 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4801 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4861 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4921 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4981 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5041 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5101 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5161 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5221 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5281 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5341 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5401 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5461 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5521 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5581 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5641 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5701 tttgataatc tcatgaccaa aatcccttaa cgtg
//
