LOCUS       sequence-341544-        8210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MCP-mCherry-GFP11-GB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8210)
  AUTHORS   Chaudhary N, Nho SH, Cho H, Gantumur N, Ra JS, Myung K, Kim H
  TITLE     Background-suppressed live visualization of genomic loci with an 
            improved CRISPR system based on a split fluorophore.
  JOURNAL   Genome Res. 2020 Sep;30(9):1306-1316. doi: 10.1101/gr.260018.119. 
            Epub 2020 Sep 4.
  PUBMED    32887690
REFERENCE   2  (bases 1 to 8210)
  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: 
            "10.1101/gr.260018.119"; journalName: "Genome Res"; date: "2020-09";
            volume: "30"; issue: "9"; pages: "1306-1316"
COMMENT     Sequence Label: pM214
FEATURES             Location/Qualifiers
     source          1..8210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2299..2602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2603..2806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2757..2777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2808..2826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2857..2881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2956..2982
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2956..2982
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2983..2994
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3391..4098
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPYDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3536..3554)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3804..3823
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4117..4137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4351..4512
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             4540..4560
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    4600..5188
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4653..4673)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5071..5082)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5263..5496
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5615..5633)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5701..5805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5806..5874,5875..6666)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6024..6043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6837..7425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7326..7345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7579..7596
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7836..7857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7872..7877,7878..7895,7896..7902)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7910..7926
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7915..7937
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7934..7950
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7934..7950
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8000..8019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagctgat gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag
     2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
     2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
     2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2941 cctaccggta tgggctaccc ctacgacgtg cccgactacg ccatcgaagg ccgccatatg
     3001 ctagccgtta aaatggcttc taactttact cagttcgttc tcgtcgacaa tggcggaact
     3061 ggcgacgtga ctgtcgcccc aagcaacttc gctaacggga tcgctgaatg gatcagctct
     3121 aactcgcgtt cacaggctta caaagtaacc tgtagcgttc gtcagagctc tgcgcagaat
     3181 cgcaaataca ccatcaaagt cgaggtgcct aaaggcgcct ggcgttcgta cttaaatatg
     3241 gaactaacca ttccaatttt cgccacgaat tccgactgcg agcttattgt taaggcaatg
     3301 caaggtctcc taaaagatgg aaacccgatt ccctcagcaa tcgcagcaaa ctccggcatc
     3361 tacgcggatc cacctgttgc tactctcgag atggtgagca agggcgagga ggataacatg
     3421 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     3481 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     3541 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     3601 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     3661 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     3721 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     3781 ggcaccaact tcccctacga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     3841 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     3901 aagctgaagg acggaggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     3961 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     4021 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     4081 atggacgagc tgtacaagac cggtatggct agcatgccca agaagaagcg caaggtgggt
     4141 tccggcgctg gcggaagccc tgggggcggg agcggtggct ctggttcttc tgctagtggc
     4201 ggctcaacat ctatggagaa gagagaccac atggtgctgc tagagtacgt gaccgccgca
     4261 ggaatcacag acgcttcagg aggcggaggt tctggtggag gcggtagtgg cggtggaggg
     4321 tcaatcgatg gtggaggtcg gaccgaagag tacaagctta tcctgaacgg taaaaccctg
     4381 aaaggtgaaa ccaccaccga agctgttgac gctgctaccg cggaaaaagt tttcaaacag
     4441 tacgctaacg acaacggtgt tgacggtgaa tggacctacg acgacgctac caaaaccttc
     4501 acggtaaccg aaggtggtgg tagcggtggt ggtactagtc ccaagaagaa gcgcaaggtg
     4561 taagcggccg ctctggctct cgagtagggt aatctagata atcaacctct ggattacaaa
     4621 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4681 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4741 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4801 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4861 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4921 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4981 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     5041 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     5101 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     5161 cggatctccc tttgggccgc ctccccgcct gagatccttt aagaccaatg acttacaagg
     5221 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5281 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5341 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5401 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5461 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     5521 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggccc gggttaatta
     5581 aggaaagggc tagatcattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     5641 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5701 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5761 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5821 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5881 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5941 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6001 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     6061 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6121 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6181 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6241 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6301 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6361 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6421 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6481 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6541 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6601 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6661 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6721 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6781 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6841 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6901 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6961 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7021 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7081 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7141 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7201 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7261 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7321 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7381 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7441 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7501 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7561 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7621 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gcaagctcat ggctgactaa
     7681 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     7741 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ccccgtggca cgacaggttt
     7801 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7861 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7921 taacaatttc acacaggaaa cagctatgac atgattacga atttcacaaa taaagcattt
     7981 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     8041 atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt
     8101 accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt
     8161 tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
