LOCUS       sequence-470717-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1428m 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: AiP15052 -
            pAAV-AiE1428m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   46..186
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   46..175
                     /label=AAV2 ITR
     misc_feature    193..220
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    221..248
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      317..772
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(404..423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     614..635
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(789..808)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     908..930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(968..986)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1054..1158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1159..1227,1228..2019)
                     /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(1377..1396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2190..2778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2679..2698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2835..2862
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2863..2890
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     repeat_region   2897..3026
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3916..3925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3922..3924,3925..3927,3928..4638)
                     /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(4228..4247)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4575..4596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4648..4704
                     /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             4717..4782
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4813..5190
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(5277..5297)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5484..5595
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
       61 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
      121 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
      181 tgcaggggcg cctatcagca cacaattgcc cattatacgc gcgtataatg gactattgtg
      241 tgctgatatg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
      301 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
      361 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
      421 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
      481 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
      541 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
      601 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
      661 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
      721 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
      781 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
      841 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
      901 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
      961 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1021 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1141 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1201 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1261 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1321 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1381 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1441 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1501 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1561 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1621 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1681 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     1741 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1801 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1861 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     1921 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1981 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2041 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2101 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2161 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2221 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2281 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2341 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2401 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2461 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2521 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2581 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2641 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2701 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2761 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2821 tggccttttg ctcatatcag cacacaattg cccattatac gcgcgtataa tggactattg
     2881 tgtgctgata acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
     2941 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     3001 ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt ctttagtagt ggctctcctt
     3061 ttgttctcca aaacactatg aaaataacta tatatggaaa ttatttaacc tccttccaaa
     3121 aaaaaccctt ataatttggt tttgaacttt gcaataatac acattgtgga aaacattcag
     3181 gatattatct ggctctagaa aagaaataaa ttaagcaaag gcgctaagta cagcaattta
     3241 cttccaatga cagtacatgc tgcctatgta gattgctttt cacaaaacac acacatttaa
     3301 tgttgtgtgt taatcagctc catgttcaat aggaaaatga ttagggcaca aacatttcat
     3361 aataggaagt tatgattgag cactgagaat gatatctcct gacttgctct tggaacagat
     3421 aatcaagttg tctttcaaag attgaacctg cagtgaatgt tttacttgag aaataaatga
     3481 aattgattat atattctagg caaaaaaata cacactataa ttactttcaa tctccttgta
     3541 acattttttc cccactagta gatagcttca caattgttag aaaaatacaa gcagatataa
     3601 gcaaaagtga gctaatctag caattagctg caggagagac cgtttgggtt aggtgatcta
     3661 tcttttcacc aagaagatgt cacaaacatc ctgttcagtc aaatttaaag acatgtgagt
     3721 gtgagccata aactccatca cttctctgaa gggcatcttg gatggattta tttgcaggat
     3781 ttcagaaaac agtatgtgct ttcaccaaga tttgcaaatc atgagctcgg gctgggcata
     3841 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
     3901 ctagcgctac cggtcgccac catggtctcc aagggcgagg agctgttcac cggggtggtg
     3961 cccatcctgg tcgagctgga cggcgatgtc aacggccaca agttcagcgt gtccggcgag
     4021 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     4081 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     4141 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4201 gtccaggagc gcaccatctt cttcaaagac gacggcaact acaagacccg cgccgaggtg
     4261 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     4321 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     4381 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     4441 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4501 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     4561 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4621 atggacgagc tgtacaaggg ctctggtgct accaacttct cactgttgaa acaggcaggg
     4681 gatgtagagg agaatccagg gcctggtgct agtggagact acaaagacca tgacggagat
     4741 tataaagatc atgacatcga ttacaaggat gacgatgaca agtccggact cagatctgga
     4801 ggctccggag gcccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag
     4861 gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc
     4921 tattccatct atgtgtataa ggttctgaag caggtccacc ctgacaccgg catttcgtcc
     4981 aaggccatgg gcatcatgaa ctcgtttgtg aacgacattt tcgagcgcat cgcaggtgag
     5041 gcttcccgcc tggcgcatta caacaagcgc tcgaccatca cctccaggga gatccagacg
     5101 gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag
     5161 gccatcacca agtacaccag cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat
     5221 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5281 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5341 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     5401 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5461 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     5521 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     5581 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     5641 gcaggacagc aagggggagg attggga
//
