LOCUS       sequence-462703-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0947m 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 5667)
  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 5667)
  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: AiP13314 -
            pAAV-AiE0947m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5667
                     /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      1133..1142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1139..1141,1142..1144,1145..1855)
                     /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(1445..1464)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1792..1813
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1865..1921
                     /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             1934..1999
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2030..2407
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2494..2514)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2701..2812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2930..3070
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2930..3059
                     /label=AAV2 ITR
     rep_origin      3145..3600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3232..3251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3442..3463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3617..3636)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3736..3758
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3796..3814)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3882..3986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3987..4055,4056..4847)
                     /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(4205..4224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5018..5606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5507..5526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttggtag agccttatga cttgcctatc cctatttgag
      181 aaggaagaat agggtggatt ccttctccag tcacctcctg tatcacaatg gaattgtact
      241 ttctattgat gagatcccta ggtgattcaa ttaaaaagca cttaacttgt gagccagaag
      301 gtccttccaa tgctggatcc actattgatt gactttaaac tatgatttct ttcctctggc
      361 ttaatttctt caccatcata ataaggccgc atgcctctca gtcacttatt gctcacagtc
      421 accactgttg gttgggttag atctccctgg tttggcttca agtcattgca gtgctcaatg
      481 tgtggctctc ccgggcatgt tccttcctga agggcagctg tgtatgaata gctaatggtt
      541 gtaatcaaag aggcagtaaa tactgctgaa tgaagatagg actgtgtcaa aggaaaggag
      601 ttattttgga aatctccatc ccaggaaatg agtagaattg tctgtttgag tcagaagtag
      661 ccccagtgat tggcgtttat ctgaagaaca acatgctgtg gtagaagtgg ccaagtgtgg
      721 gctgcctcag aaaaaacggg cagccttaga acagtcagcc caagagccac ctggagccat
      781 ttcagccctg attcccatga cctctatgac attggcaact tcacatccag aaactgcgac
      841 ttatctatct gaaaactggt cttcaaagaa agcagttggt caaagggatc ccaggagctc
      901 aataatatgg catggtcatg aaattttaaa taacacatat ttataagggt gtcagtgccc
      961 tgaagagttc cccttggagg aaattttaat accaactccc aactaccctg agaaaaatca
     1021 ctcaatgcct aagagctcag agctcgggct gggcataaaa gtcagggcag agccatctat
     1081 tgcttacatt tgcttctggg atccagatct ttcgaagcta gcgctaccgg tcgccaccat
     1141 ggtctccaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1201 cgatgtcaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1261 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1321 cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     1381 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1441 caaagacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1501 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1561 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     1621 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     1681 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1741 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1801 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggctc
     1861 tggtgctacc aacttctcac tgttgaaaca ggcaggggat gtagaggaga atccagggcc
     1921 tggtgctagt ggagactaca aagaccatga cggagattat aaagatcatg acatcgatta
     1981 caaggatgac gatgacaagt ccggactcag atctggaggc tccggaggcc cagagccagc
     2041 gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg cgcagaagaa
     2101 aggcggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg tgtacaaggt
     2161 tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca tcatgaattc
     2221 gtttgtgaac gacattttcg agcgcatcgc aggagaggct tcccgcctgg cgcattacaa
     2281 caagcgctcg accatcacct cccgggagat ccagacggcc gtgcgcctgc tgctgcctgg
     2341 ggagttggcc aagcacgccg tgtccgaggg tactaaggcc atcaccaagt acaccagcgc
     2401 taagtaatga ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa
     2461 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2521 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2581 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     2641 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga
     2701 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2761 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2821 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2881 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     2941 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3001 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3061 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3121 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3181 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     3241 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3301 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3361 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3421 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     3481 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     3541 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     3601 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     3661 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3721 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3781 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3841 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3901 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3961 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4021 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4081 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4141 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4201 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4261 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4321 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4381 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4441 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4501 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4561 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4621 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4681 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4741 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4801 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4861 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4921 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4981 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5041 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5101 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5161 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5221 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5281 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5341 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5401 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5461 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5521 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5581 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5641 ccttttgctg gccttttgct cacatgt
//
