LOCUS       sequence-229037-        3075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector for cloning LbCpf1-spCas9 MS2-fgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3075)
  AUTHORS   Kweon J, Jang AH, Kim DE, Yang JW, Yoon M, Rim Shin H, Kim JS, Kim Y
  TITLE     Publisher Correction: Fusion guide RNAs for orthogonal gene 
            manipulation with Cas9 and Cpf1.
  JOURNAL   Nat Commun. 2018 Jan 16;9(1):303. doi: 10.1038/s41467-017-02686-8.
  PUBMED    29335559
REFERENCE   2  (bases 1 to 3075)
  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: "Nat 
            Commun."; date: "2018-01-16"; volume: "9(1):303. doi"; pages: " 
            10.1038/s41467-017-02686-8"
COMMENT     Sequence Label: pFGRNA-LbCpf1-spCas9-MS2
FEATURES             Location/Qualifiers
     source          1..3075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     344..362
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(400..422)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     522..541
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     735..757
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     749..766
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     750..766
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        812..830
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        882..900
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(1066..1083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1237..1825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1317..1336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1996..2787,2788..2856))
                     /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     2619..2638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2857..2961)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg aatttctact aagtgtagat tgagaccaca tttccccgaa aagtgccacc
      301 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      361 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      421 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      481 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      541 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
      601 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      661 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
      721 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttgcatg
      781 cctgcagggt ctctgtttta gagctaggcc aacatgagga tcacccatgt ctgcagggcc
      841 tagcaagtta aaataaggct agtccgttat caacttggcc aacatgagga tcacccatgt
      901 ctgcagggcc aagtggcacc gagtcggtgc ttttttttct agattcgcga tgtacgggcc
      961 agatatacgc gttgacattg attattgact agttgtcttc ctgcattaat gaatcggcca
     1021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2101 cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac cggctccaga
     2161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2281 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2581 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2881 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2941 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacgaaga
     3001 ctcaattgtc gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg
     3061 gccgccccct tcacc
//
