LOCUS       sequence-209271-        7421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing AsCpf1 (RR variant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7421)
  AUTHORS   Tekin H, Simmons S, Cummings B, Gao L, Adiconis X, Hession CC, 
            Ghoshal A, Dionne D, Choudhury SR, Yesilyurt V, Sanjana NE, Shi X, 
            Lu C, Heidenreich M, Pan JQ, Levin JZ, Zhang F
  TITLE     Effects of 3D culturing conditions on the transcriptomic profile of 
            stem-cell-derived neurons.
  JOURNAL   Nat Biomed Eng. 2018 Jul;2(7):540-554. doi: 
            10.1038/s41551-018-0219-9. Epub 2018 Apr 9.
  PUBMED    30271673
REFERENCE   2  (bases 1 to 7421)
  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"; doi: 
            "10.1038/s41551-018-0219-9"; journalName: "Nat Biomed Eng"; date: 
            "2018-07"; volume: "2"; issue: "7"; pages: "540-554"
COMMENT     Sequence Label: hSyn1-AsCpf1(RR)
FEATURES             Location/Qualifiers
     source          1..7421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        164..611
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             652..678
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             682..702
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             712..4629
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPRCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      4349..4358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4639..4687
                     /note="synthetic polyadenylation signal"
     repeat_region   4695..4835
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4695..4824
                     /label=AAV2 ITR
     rep_origin      4910..5365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4997..5016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5207..5228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5382..5401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5501..5523
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5561..5579)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5647..5751
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5752..5820,5821..6612)
                     /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(5970..5989)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6783..7371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7272..7291
                     /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 gcggcctcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
      181 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      241 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      301 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      361 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      421 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      481 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      541 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      601 tgagagcgca gtcgagaagg taccggatcc ccgggtaccg gtgccaccat gtacccatac
      661 gatgttccag attacgcttc gccgaagaaa aagcgcaagg tcgaagcgtc cacacagttc
      721 gagggcttta ccaacctgta tcaggtgagc aagacactgc ggtttgagct gatcccacag
      781 ggcaagaccc tgaagcacat ccaggagcag ggcttcatcg aggaggacaa ggcccgcaat
      841 gatcactaca aggagctgaa gcccatcatc gatcggatct acaagaccta tgccgaccag
      901 tgcctgcagc tggtgcagct ggattgggag aacctgagcg ccgccatcga ctcctataga
      961 aaggagaaaa ccgaggagac aaggaacgcc ctgatcgagg agcaggccac atatcgcaat
     1021 gccatccacg actacttcat cggccggaca gacaacctga ccgatgccat caataagaga
     1081 cacgccgaga tctacaaggg cctgttcaag gccgagctgt ttaatggcaa ggtgctgaag
     1141 cagctgggca ccgtgaccac aaccgagcac gagaacgccc tgctgcggag cttcgacaag
     1201 tttacaacct acttctccgg cttttatgag aacaggaaga acgtgttcag cgccgaggat
     1261 atcagcacag ccatcccaca ccgcatcgtg caggacaact tccccaagtt taaggagaat
     1321 tgtcacatct tcacacgcct gatcaccgcc gtgcccagcc tgcgggagca ctttgagaac
     1381 gtgaagaagg ccatcggcat cttcgtgagc acctccatcg aggaggtgtt ttccttccct
     1441 ttttataacc agctgctgac acagacccag atcgacctgt ataaccagct gctgggagga
     1501 atctctcggg aggcaggcac cgagaagatc aagggcctga acgaggtgct gaatctggcc
     1561 atccagaaga atgatgagac agcccacatc atcgcctccc tgccacacag attcatcccc
     1621 ctgtttaagc agatcctgtc cgataggaac accctgtctt tcatcctgga ggagtttaag
     1681 agcgacgagg aagtgatcca gtccttctgc aagtacaaga cactgctgag aaacgagaac
     1741 gtgctggaga cagccgaggc cctgtttaac gagctgaaca gcatcgacct gacacacatc
     1801 ttcatcagcc acaagaagct ggagacaatc agcagcgccc tgtgcgacca ctgggataca
     1861 ctgaggaatg ccctgtatga gcggagaatc tccgagctga caggcaagat caccaagtct
     1921 gccaaggaga aggtgcagcg cagcctgaag cacgaggata tcaacctgca ggagatcatc
     1981 tctgccgcag gcaaggagct gagcgaggcc ttcaagcaga aaaccagcga gatcctgtcc
     2041 cacgcacacg ccgccctgga tcagccactg cctacaaccc tgaagaagca ggaggagaag
     2101 gagatcctga agtctcagct ggacagcctg ctgggcctgt accacctgct ggactggttt
     2161 gccgtggatg agtccaacga ggtggacccc gagttctctg cccggctgac cggcatcaag
     2221 ctggagatgg agccttctct gagcttctac aacaaggcca gaaattatgc caccaagaag
     2281 ccctactccg tggagaagtt caagctgaac tttcagatgc ctacactggc ccggggctgg
     2341 gacgtgaata aggagaagaa caatggcgcc atcctgtttg tgaagaacgg cctgtactat
     2401 ctgggcatca tgccaaagca gaagggcagg tataaggccc tcagcttcga gcccacagag
     2461 aaaaccagcg agggctttga taagatgtac tatgactact ttccggatgc cgccaagatg
     2521 atcccacggt gcagcaccca gctgaaggcg gtgaccgccc actttcagac ccacacaacc
     2581 cccatcctgc tgtccaacaa tttcatcgag cctctggaga tcacaaagga gatctacgac
     2641 ctgaacaatc ctgagaagga gccaaagaag tttcagacag cgtacgccaa gaaaaccggc
     2701 gaccagaagg gctacagaga ggccctgtgc aagtggatcg acttcacaag ggattttctg
     2761 tccaagtata ccaagacaac ctctatcgat ctgtctagcc tgcggccatc ctctcagtat
     2821 aaggacctgg gcgagtacta tgccgagctg aatcccctgc tgtaccacat cagcttccag
     2881 agaatcgccg agaaggagat catggatgcc gtggagacag gcaagctgta cctgttccag
     2941 atctataaca aggactttgc caagggccac cacggcaagc ctaatctgca cacactgtat
     3001 tggaccggtc tgttttctcc agagaacctg gccaagacaa gcatcaagct gaatggccag
     3061 gccgagctgt tctaccgccc taagtccagg atgaagagga tggcacaccg gctgggagag
     3121 aagatgctga acaagaagct gaaggatcag aaaaccccaa tccccgacac cctgtaccag
     3181 gagctgtacg actatgtgaa tcacagactg tcccacgacc tgtctgatga ggccagggcc
     3241 ctgctgccca acgtgatcac caaggaggtg tctcacgaga tcatcaagga taggcgcttt
     3301 accagcgaca agttcttttt ccacgtgcct atcacactga actatcaggc cgccaattcc
     3361 ccatctaagt tcaaccagag ggtgaatgcc tacctgaagg agcaccccga gacacctatc
     3421 atcggcatcg atcggggcga gagaaacctg atctatatca cagtgatcga ctccaccggc
     3481 aagatcctgg agcagcggag cctgaacacc atccagcagt ttgattacca gaagaagctg
     3541 gacaacaggg agaaggagag ggtggcagca aggcaggcct ggtctgtggt gggcacaatc
     3601 aaggatctga agcagggcta tctgagccag gtcatccacg agatcgtgga cctgatgatc
     3661 cactaccagg ccgtggtggt gctggagaac ctgaatttcg gctttaagag caagaggacc
     3721 ggcatcgccg agaaggccgt gtaccagcag ttcgagaaga tgctgatcga taagctgaat
     3781 tgcctggtgc tgaaggacta tccagcagag aaagtgggag gcgtgctgaa cccataccag
     3841 ctgacagacc agttcacctc ctttgccaag atgggcaccc agtctggctt cctgttttac
     3901 gtgcctgccc catatacatc taagatcgat cccctgaccg gcttcgtgga ccccttcgtg
     3961 tggaaaacca tcaagaatca cgagagccgc aagcacttcc tggagggctt cgactttctg
     4021 cactacgacg tgaaaaccgg cgacttcatc ctgcacttta agatgaacag aaatctgtcc
     4081 ttccagaggg gcctgcccgg ctttatgcct gcatgggata tcgtgttcga gaagaacgag
     4141 acacagtttg acgccaaggg cacccctttc atcgccggca agagaatcgt gccagtgatc
     4201 gagaatcaca gattcaccgg cagataccgg gacctgtatc ctgccaacga gctgatcgcc
     4261 ctgctggagg agaagggcat cgtgttcagg gatggctcca acatcctgcc aaagctgctg
     4321 gagaatgacg attctcacgc catcgacacc atggtggccc tgatccgcag cgtgctgcag
     4381 atgcggaact ccaatgccgc cacaggcgag gactatatca acagccccgt gcgcgatctg
     4441 aatggcgtgt gcttcgactc ccggtttcag aacccagagt ggcccatgga cgccgatgcc
     4501 aatggcgcct accacatcgc cctgaagggc cagctgctgc tgaatcacct gaaggagagc
     4561 aaggatctga agctgcagaa cggcatctcc aatcaggact ggctggccta catccaggag
     4621 ctgcgcaact aagaattcaa taaaagatct ttattttcat tagatctgtg tgttggtttt
     4681 ttgtgtgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4741 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     4801 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     4861 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     4921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4981 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5101 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     5161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5221 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     5281 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5341 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     5401 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5461 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5521 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5581 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5641 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5701 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5761 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5821 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5881 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5941 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6001 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6061 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6121 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6181 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6241 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6301 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6361 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6421 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc
     6481 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6541 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6601 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6661 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6721 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6781 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6841 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6901 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6961 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7021 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7081 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7141 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7201 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7261 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7321 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7381 aacgcggcct ttttacggtt cctggccttt tgctcacatg t
//
