LOCUS       sequence-305645-        4359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rfx Cas13d guide cloning pUC19 vector with constitutively expressed 
            tagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4359)
  AUTHORS   Abbott TR, Dhamdhere G, Liu Y, Lin X, Goudy L, Zeng L, Chemparathy 
            A, Chmura S, Heaton NS, Debs R, Pande T, Endy D, La Russa MF, Lewis 
            DB, Qi LS
  TITLE     Development of CRISPR as an Antiviral Strategy to Combat SARS-CoV-2 
            and Influenza.
  JOURNAL   Cell. 2020 May 14;181(4):865-876.e12. doi: 
            10.1016/j.cell.2020.04.020. Epub 2020 Apr 29.
  PUBMED    32353252
REFERENCE   2  (bases 1 to 4359)
  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: 
            "10.1016/j.cell.2020.04.020"; journalName: "Cell"; date: 
            "2020-05-14- 14"; volume: "181"; issue: "4"; pages: "865-876.e12"
COMMENT     Sequence Label: pSLQ5429_pUC_hU6-crScaffold_EF1a-BFP
FEATURES             Location/Qualifiers
     source          1..4359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..255
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7..27
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     178..197
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        257..292
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        311..346
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     promoter        401..1579
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          632..1570
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1527..1547
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(2322..2339)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2328..2552
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2579..2683
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2684..2752,2753..3544)
                     /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(2902..2921)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3715..4303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4204..4223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctagcgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
       61 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
      121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
      181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      241 aaaggacgaa acaccgcaag taaaccccta ccaactggtc ggggtttgaa acgggtcttc
      301 gagaagacct caagtaaacc cctaccaact ggtcggggtt tgaaactttt tttttcacct
      361 atgtatcgta tcaagtacta gaactcccca gtgggaattc ggctccggtg cccgtcagtg
      421 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      481 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      541 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
      601 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
      661 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
      721 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
      781 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
      841 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
      901 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
      961 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     1021 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     1081 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     1141 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     1201 gccgcttccc ggccctgctg cagggagctc aaaatgaagg acgcggcgct cgggagagcg
     1261 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1321 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     1381 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1441 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1501 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     1561 tccatttcag gtgtcgtgac gtacggccac catgagcatg tccgaactga ttaaagagaa
     1621 catgcacatg aaactttata tggaaggtac ggtcgacaac caccatttca aatgtacctc
     1681 cgaaggagag ggcaaaccct acgagggcac ccaaacgatg agaattaaag tagtagaagg
     1741 tgggccgctg ccctttgcct ttgacattct ggcaacttct tttctctacg ggagtaaaac
     1801 ttttatcaac catacgcaag gcattccaga cttctttaag cagtcatttc ccgaggggtt
     1861 tacttgggag cgggtcacaa catacgagga cggcggcgtg ctgaccgcta ctcaggacac
     1921 cagcctgcaa gacgggtgtt tgatctacaa tgtgaagatt cgaggcgtaa actttacttc
     1981 aaacggccca gttatgcaga aaaaaaccct cggctgggag gcttttacag agactcttta
     2041 cccagccgac ggtggtttgg aaggccgaaa cgacatggca ttgaaattgg ttggcgggag
     2101 ccaccttata gcgaatataa agacgaccta tcggtctaaa aagcccgcca aaaatttgaa
     2161 gatgcctggt gtatattacg tcgactatag gcttgagaga ataaaagaag ctaacaacga
     2221 aacctatgtg gagcaacatg aggttgcagt tgcacgctac tgcgatctgc cgagcaaatt
     2281 ggggcacaag ctgaattgat cagctagagc tcgctaatca gcctcgactg tgccttctag
     2341 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2401 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2461 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagataatag
     2521 caggcatgct ggggatgcgg tgggctctat ggtctagatt tagggtaggg gaaatgtgcg
     2581 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2641 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2701 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2761 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2821 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2881 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2941 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3001 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3061 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3121 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3181 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3241 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3301 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3361 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     3421 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3481 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3541 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3601 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3661 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3721 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3781 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3841 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3901 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3961 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4021 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4081 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4141 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4201 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4261 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4321 ctttttacgg ttcctggcct tttgctggcc ttttgctca
//
