LOCUS       sequence-194498-        7802 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes dCas9(N) fused to CXCR4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7802)
  AUTHORS   Baeumler TA, Ahmed AA, Fulga TA
  TITLE     Engineering Synthetic Signaling Pathways with Programmable 
            dCas9-Based Chimeric Receptors.
  JOURNAL   Cell Rep. 2017 Sep 12;20(11):2639-2653. doi: 
            10.1016/j.celrep.2017.08.044.
  PUBMED    28903044
REFERENCE   2  (bases 1 to 7802)
  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.celrep.2017.08"; journalName: "Cell Rep"; date: 
            "2017-09-12- 12"; volume: "20"; issue: "11"; pages: "2639-2653"
COMMENT     Sequence Label: pCXCR4_TCS(Q'L)_HA-dCas9(N)_P2A-Puro-WPRE
FEATURES             Location/Qualifiers
     source          1..7802
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1499..1525
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1541..1564,1565..1567,1568..3142)
                     /codon_start=1
                     /product="N-terminal portion of catalytically inactive 
                     Streptococcus pyogenes dCas9 carrying the D10A mutation 
                     (Zetsche et al., 2015)"
                     /label=dCas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMR"
     CDS             3173..3229
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(3236..3255)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3732..3752
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3869..3889)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4287..4298)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4429..4446)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4435..4642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4651..4791
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4651..4780
                     /label=AAV2 ITR
     rep_origin      4866..5321
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4953..4972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5163..5184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5338..5357)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5457..5479
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5517..5535)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5603..5707
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5708..5776,5777..6568)
                     /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(5926..5945)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6739..7327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7228..7247
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7398..7683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
       61 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      121 tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt aatgtttaat
      181 tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcg ccaccatgaa
      241 gacgatcatc gccctgagct acatcttctg cctggtattc gccgactaca aggacgatga
      301 tgacgccagc atcgatatgg aggggatatc aatctacaca tcagataatt acacggaaga
      361 gatgggttcc ggcgattacg actctatgaa agagccgtgt tttagagagg aaaacgcgaa
      421 ctttaacaag atctttctcc ccaccatcta cagcatcatc tttctcacag gcatcgtagg
      481 gaacggcctg gtcatcctgg tgatgggata ccaaaagaag ctgaggtcaa tgaccgacaa
      541 gtataggctc catctgtccg tggccgacct cctgtttgtg attaccctgc ctttttgggc
      601 agttgacgct gtcgctaatt ggtacttcgg caacttcctc tgtaaggcag tgcacgttat
      661 ctacactgtg aatctttata gttccgtctt gatcttggcc tttatcagtc tcgacaggta
      721 tttggcgatt gtgcacgcta ccaactcaca acgacctaga aaactcctgg ctgagaaggt
      781 ggtttacgtt ggtgtgtgga ttccagctct cctgttgaca ataccagact tcattttcgc
      841 taatgtgagc gaggccgatg acagatacat ttgtgaccga ttttacccaa acgatctgtg
      901 ggtagtggta tttcagttcc aacacattat ggtcgggctg atcttgcccg gcattgtcat
      961 actgtcttgc tactgcatca ttatttctaa gctgtcacac tcaaaaggcc accaaaagag
     1021 gaaggctctg aaaacaacgg tgatcctgat actggccttc ttcgcatgtt ggctgcccta
     1081 ctatatcggc atcagcattg actcatttat actcctggaa attatcaagc agggctgcga
     1141 gttcgagaac accgttcata agtggatttc tataaccgag gccctcgcct tctttcactg
     1201 ttgtttgaat ccgattctct acgcgtttct tggcgccaaa tttaaaacaa gcgcccaaca
     1261 tgcactgaca tcagtgtcta gggggagctc tctgaaaatc ctctccaagg gaaaacgagg
     1321 cggacatagc agtgtcagca ctgagtccga atccagctca tttcatagct ctatcgatac
     1381 cggtggacgc accccaccca gcctgggtcc ccaagatgag tcctgcacca ccgccagctc
     1441 ctccctggcc aaggacactt catcgaccgg tgagaacctg tacttccagc taactagtta
     1501 cccatacgac gtaccagatt acgctggagg ttctggatcc gacaagaagt acagcatcgg
     1561 cctggccatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
     1621 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
     1681 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
     1741 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
     1801 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
     1861 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
     1921 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
     1981 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     2041 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     2101 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     2161 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     2221 gctgcccggc gagaagaaga atggcctgtt cggcaacctg attgccctga gcctgggcct
     2281 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     2341 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     2401 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     2461 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     2521 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     2581 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     2641 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     2701 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     2761 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     2821 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     2881 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     2941 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     3001 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     3061 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     3121 atacgtgacc gagggaatga gaggtggcgg tggctcgggt accggatctg gagccaccaa
     3181 tttttctctg ctgaagcagg ctggtgatgt tgaagaaaac cctggacctg aattcatgac
     3241 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3301 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3361 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3421 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3481 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3541 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3601 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3661 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3721 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3781 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aaggggtaac taagtaagga
     3841 tcgatccaag atatcgtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3901 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3961 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4021 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4081 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4141 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4201 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     4261 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4321 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4381 tctccctttg ggccgcctcc ccgccaattc ctagagctcg ctgatcagcc tcgactgtgc
     4441 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4501 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4561 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4621 agaatagcag gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc
     4681 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4741 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     4801 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     4861 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4921 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     4981 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5041 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5101 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5161 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     5221 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     5281 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5341 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5401 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5461 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5521 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5581 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5641 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5701 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5761 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5821 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5881 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5941 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6001 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6061 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6121 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6181 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6241 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6301 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6361 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6421 tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6481 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6541 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6601 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6661 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6721 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6781 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6841 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6901 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6961 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7021 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7081 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7141 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7201 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7261 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7321 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7381 tcacatctag aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     7441 aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa
     7501 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7561 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac
     7621 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7681 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
     7741 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
     7801 gg
//
