LOCUS       sequence-384988-        7453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression foldon-GFP-PCP protein for imaging of nonrepetitive DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7453)
  AUTHORS   Lyu XY, Deng Y, Huang XY, Li ZZ, Fang GQ, Yang D, Wang FL, Kang W, 
            Shen EZ, Song CQ
  TITLE     CRISPR FISHer enables high-sensitivity imaging of nonrepetitive DNA 
            in living cells through phase separation-mediated signal 
            amplification.
  JOURNAL   Cell Res. 2022 Nov;32(11):969-981. doi: 10.1038/s41422-022-00712-z. 
            Epub 2022 Sep 14.
  PUBMED    36104507
REFERENCE   2  (bases 1 to 7453)
  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: 
            "10.1038/s41422-022-00712-z"; journalName: "Cell Res"; date: 
            "2022-11"; volume: "32"; issue: "11"; pages: "969-981"
COMMENT     Sequence Label: pHAGE-EFS-foldon-GFP- PCP
FEATURES             Location/Qualifiers
     source          1..7453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    643..664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(679..684,685..702,703..709)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    717..733
                     /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     722..744
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     741..757
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     741..757
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     807..826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             1018..1651
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1698..1823
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2320..2553
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2738..2782
                     /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    3075..3192
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3282..3493
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3539..3548
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3677..4387
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4829..4849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    4860..5448
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4913..4933)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5331..5342)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5523..5756
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5875..5893)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5961..6065
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6066..6134,6135..6926)
                     /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(6284..6303)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7097..7453,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagca agctcatggc
      481 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
      541 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc cgtggcacga
      601 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      661 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      721 gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt tcacaaataa
      781 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      841 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
      901 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
      961 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagttgg
     1021 aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac
     1081 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
     1141 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
     1201 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     1261 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
     1321 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
     1381 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
     1441 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
     1501 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     1561 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     1621 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     1681 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     1741 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     1801 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     1861 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     1921 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     1981 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     2041 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     2101 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     2161 cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     2221 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     2281 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     2341 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     2401 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     2461 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     2521 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     2581 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     2641 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     2701 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     2761 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     2821 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     2881 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     2941 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     3001 agacagatcc attcgattag tgaacggatc tcgacggtat cgccgaattg acaaatggca
     3061 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     3121 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     3181 attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggac tagcgatgct
     3241 agctgatgcg ggcccttcga gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     3301 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     3361 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     3421 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     3481 cgccagaaca caggtgtcgt gacgcggttc cggttaaggt gaacgcgtgc aggctggcgc
     3541 caccatgggc tacatccccg aggcccccag ggacggccag gcctacgtga ggaaggacgg
     3601 cgagtgggtg ctgctgagca ccttcctgag tggcggagga ggttcaggag gcggcggaag
     3661 cggtggagga ggttctcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt
     3721 ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga
     3781 cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc
     3841 ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga
     3901 ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg
     3961 cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg
     4021 cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat
     4081 cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa
     4141 acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt
     4201 gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc
     4261 agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga
     4321 tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact
     4381 gtacaaaatg ggttccaaaa ccatcgttct ttcggtcggc gaggctactc gcactctgac
     4441 tgagatccag tccaccgcag accgtcagat cttcgaagag aaggtcgggc ctctggtggg
     4501 tcggctgcgc ctcacggctt cgctccgtca aaacggagcc aagaccgcgt atcgcgtcaa
     4561 cctaaaactg gatcaggcgg acgtcgttga ttccggactt ccgaaagtgc gctacactca
     4621 ggtatggtcg cacgacgtga caatcgttgc gaatagcacc gaggcctcgc gcaaatcgtt
     4681 gtacgatttg accaagtccc tcgtcgcgac ctcgcaggtc gaagatcttg tcgtcaacct
     4741 tgtgccgctg ggccgtggtg gcggagggac tagtggaggt ggatccggaa gttctggcgg
     4801 ccgctctggc atcgatggga gcctcgagcc aaagaaaaag cggaaagtgt aatctagata
     4861 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4921 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4981 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5041 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5101 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5161 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5221 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5281 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5341 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5401 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct gagatccttt
     5461 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     5521 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     5581 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5641 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5701 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     5761 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     5821 tgagaggccc gggttaatta aggaaagggc tagatcattc ttgaagacga aagggcctcg
     5881 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5941 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6001 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6061 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6121 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6181 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6241 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6301 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6361 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6421 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6481 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6541 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6601 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6661 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6721 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6781 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6841 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6901 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6961 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7021 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7081 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7141 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7201 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7441 gcttggagcg aac
//
