LOCUS       sequence-331084-        3089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6-driven expression of human VEGFA targeting array presgRNA 
            compatible with CasRx..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3089)
  AUTHORS   Chuang YF, Wang PY, Kumar S, Lama S, Lin FL, Liu GS
  TITLE     Methods for in vitro CRISPR/CasRx-Mediated RNA Editing.
  JOURNAL   Front Cell Dev Biol. 2021 Jun 11;9:667879. doi: 
            10.3389/fcell.2021.667879. eCollection 2021.
  PUBMED    34178991
REFERENCE   2  (bases 1 to 3089)
  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: "Front Cell 
            Dev Biol."; date: "2021-06-11"; volume: "9:667879. doi"; pages: " 
            10.3389/fcell.2021.667879. eCollection 2021"
COMMENT     Sequence Label: pM117: VEGFA array presgRNA
FEATURES             Location/Qualifiers
     source          1..3089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..283
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     35..55
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     206..225
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        285..320
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        351..386
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        417..452
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        483..518
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     rep_origin      600..1055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(687..706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     897..918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1072..1091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1191..1213
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1251..1269)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1337..1441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1442..1510,1511..2302)
                     /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(1660..1679)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2473..3061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2962..2981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctggcctt ttgctggcct tttgctcagc tagcgagggc ctatttccca tgattccttc
       61 atatttgcat atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa
      121 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
      181 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
      241 cgatttcttg gctttatata tcttgtggaa aggacgaaac accgcaagta aacccctacc
      301 aactggtcgg ggtttgaaac tatgtgctgg ccttggtgag gtttgatccg caagtaaacc
      361 cctaccaact ggtcggggtt tgaaaccact tcgtgatgat tctgccctcc tccttccaag
      421 taaaccccta ccaactggtc ggggtttgaa actctgcatt cacatttgtt gtgctgtagg
      481 aacaagtaaa cccctaccaa ctggtcgggg tttgaaactt tttttgaatt ctgatgcggt
      541 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
      601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
      661 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
      781 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
      841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
      901 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
      961 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1021 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     1081 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     1141 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     1201 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     1261 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     1321 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1381 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1441 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1501 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1561 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1621 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1681 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1741 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1801 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1861 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1921 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1981 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2041 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2101 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag
     2161 ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2221 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2281 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2341 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2401 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2461 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2521 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2581 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2641 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2701 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2761 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2821 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2881 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2941 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3001 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3061 aaacgccagc aacgcggcct ttttacggt
//
