LOCUS       sequence-199009-        4772 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression construct for dCas9 fused to GFP-binding Protein 
            (N-terminally) and mRFP (C-terminus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4772)
  AUTHORS   Anton T, Bultmann S
  TITLE     Site-specific recruitment of epigenetic factors with a modular 
            CRISPR/Cas system.
  JOURNAL   Nucleus. 2017 May 4;8(3):279-286. doi: 
            10.1080/19491034.2017.1292194. Epub 2017 Feb 23.
  PUBMED    28448738
REFERENCE   2  (bases 1 to 4772)
  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: 
            "10.1080/19491034.2017.1292194"; journalName: "Nucleus"; date: 
            "2017-05-4- 4"; volume: "8"; issue: "3"; pages: "279-286"
COMMENT     Sequence Label: pCAG-GBP-dCas9-mRFP
FEATURES             Location/Qualifiers
     source          1..4772
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    47..509
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(103..120)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     330..349
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      514..523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             520..918
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    1076..1197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1113..1132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1167..1186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1245..1380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1486..1503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1657..2245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1737..1756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2416..3207,3208..3276))
                     /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     3039..3058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3277..3381)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3449..3467
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3505..3527)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3627..3646
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3713..4168)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3850..3871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4062..4081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4294..4316
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4308..4325
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4309..4325
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4332..4350
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        4419..4722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
       61 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
      121 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
      181 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
      241 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
      301 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
      361 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
      421 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
      481 tggttttcct ttgaaaaaca cgatgataag cttgccacca tggccaagcc tttgtctcaa
      541 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
      601 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
      661 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
      721 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
      781 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
      841 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
      901 tatgtgtggg agggctaagc acttcgtggc cgaggagcag gactgacacg tgctacgaga
      961 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     1021 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     1081 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1141 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     1201 tgtctgtata ccgtcgacct cgaggagctt tttgcaaaag cctaggcctc caaaaaagcc
     1261 tcttcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     1321 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     1381 gggcggagtt aggggcggga ctatggttgc tgactaattg ctgcattaat gaatcggcca
     1441 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1501 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1561 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1621 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1681 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1741 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1801 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1861 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1921 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1981 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2041 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2101 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2161 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2281 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2341 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2401 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2461 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2521 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2581 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2641 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2701 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2761 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2821 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2881 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2941 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3001 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3061 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3121 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3181 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3241 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3301 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3361 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3421 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     3481 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3541 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3601 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     3661 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     3721 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3781 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3841 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3901 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3961 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4021 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4081 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4141 gcgctcaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg
     4201 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     4261 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     4321 ccagtgaatt gtaatacgac tcactatagg acgcgtccta gttattaata gtaatcaatt
     4381 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4441 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4501 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4561 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     4621 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     4681 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     4741 ttctgcttca ctctccccat ctcccccccc tc
//
