LOCUS       sequence-461888-        5381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0655m 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 5381)
  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 5381)
  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: AiP13657 -
            pAAV-AiE0655m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5381
                     /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      847..856
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(853..855,856..858,859..1569)
                     /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(1159..1178)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1506..1527
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1579..1635
                     /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             1648..1713
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1744..2121
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2208..2228)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2415..2526
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2644..2784
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2644..2773
                     /label=AAV2 ITR
     rep_origin      2859..3314
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2946..2965)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3156..3177
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3331..3350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3450..3472
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3510..3528)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3596..3700
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3701..3769,3770..4561)
                     /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(3919..3938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4732..5320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5221..5240
                     /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 gcgtcacaag gtgaggaact gtgttaaagg gtcgcagcat
      181 taggaaggtt gagaaccact ggtctagatg gaactggggc agggcaagcc aggcgagatg
      241 gataaacatg tctctactcc cccccccccc cccccagacg ttgttgacta agctgtgtgg
      301 gcaggaccgg ctcctgaggc agctacagga ggatctggac aagaggcagg aggagaaggt
      361 acagaagggg tgatgtgcct tagctgctcc tgcccttgat tcctccccag gcaccagggt
      421 cgagatgctt cccctgtggt gggcttgggg ggctcgcttg ccctcttggc ttcagaagtc
      481 caatcgctgt ggtctgtgca ggagcagctg gaggcagctt tggagctgac ccggcagcag
      541 ctgggccaag ctaccaggga ggctgcagct tccgggaagg cctggggtcg ccagcgcctt
      601 ctgcaggacc ggctggtgaa cgtgagggct gctctgtgtc acctggctca ggtgagcatc
      661 tgggaagaga ctcccaggct tgtgcaggtc actgcagaga ttccgtaggc ctgtgggttg
      721 ttctcgaaat gccagtgcag tcacaaaacc cacgagctcg ggctgggcat aaaagtcagg
      781 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
      841 ccggtcgcca ccatggtctc caagggcgag gagctgttca ccggggtggt gcccatcctg
      901 gtcgagctgg acggcgatgt caacggccac aagttcagcg tgtccggcga gggcgagggc
      961 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     1021 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     1081 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1141 cgcaccatct tcttcaaaga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1201 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1261 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1321 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1381 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1441 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1501 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1561 ctgtacaagg gctctggtgc taccaacttc tcactgttga aacaggcagg ggatgtagag
     1621 gagaatccag ggcctggtgc tagtggagac tacaaagacc atgacggaga ttataaagat
     1681 catgacatcg attacaagga tgacgatgac aagtccggac tcagatctgg aggctccgga
     1741 ggcccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact
     1801 aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag ctattccatc
     1861 tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc caaggccatg
     1921 ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggaga ggcttcccgc
     1981 ctggcgcatt acaacaagcg ctcgaccatc acctcccggg agatccagac ggccgtgcgc
     2041 ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa ggccatcacc
     2101 aagtacacca gcgctaagta atgaggcgcg ccgcggccgc gaattcgata tcataatcaa
     2161 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2221 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2281 ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc
     2341 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggct
     2401 cgagagatct tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2461 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2521 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2581 caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg accgagcggc
     2641 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2701 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2761 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2821 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     2881 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     2941 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3001 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3061 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3121 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3181 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3241 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3301 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3361 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3421 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3481 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3541 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3601 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3661 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3721 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3781 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3841 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3901 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3961 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4021 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4081 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4141 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4201 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4261 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4321 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4381 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4441 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4501 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4561 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4621 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4681 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4741 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4801 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4861 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4921 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4981 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5041 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5101 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5161 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5221 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5281 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5341 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
