LOCUS       sequence-497478-        8544 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  Mammalian lentiviral vector for the expression of the 
            Sapphire-tagged nuclear localizing 12nm nanoGEMs under the UBC 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8544)
  TITLE     Herpes simplex virus 1 fluidizes the nucleus enabling condensate 
            formation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8544)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pUBC-SV40NLS-HoTag3-Shunzhi12nm-TSapphire
FEATURES             Location/Qualifiers
     source          1..8544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(122..138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(122..138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(135..157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    146..162
                     /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)."
     promoter        complement(join(170..176,177..194,195..200))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    215..236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(353..370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(524..1112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(604..623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1283..2074,2075..2143))
                     /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     1906..1925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2144..2248)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2296..2430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2333..2352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2387..2406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             2598..3231
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3278..3403
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3900..4133
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4318..4362
                     /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    4625..4740
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        4786..5997
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5834..5853
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6019..6039
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    7110..7698
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7163..7183)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7581..7592)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7905..8538
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
       61 agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg gcgtaatcat
      121 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      181 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      241 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      301 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
      361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
      481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      961 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1081 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1141 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1201 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1261 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1321 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1381 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1441 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1501 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1561 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     1621 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1681 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1741 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1801 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1861 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1921 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1981 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2041 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2101 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     2281 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2401 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
     2461 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
     2521 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
     2581 actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga
     2641 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     2701 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     2761 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     2821 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     2881 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     2941 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     3001 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     3061 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     3121 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     3181 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     3241 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     3301 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     3361 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     3421 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     3481 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     3541 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     3601 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     3661 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     3721 taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg
     3781 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     3841 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     3901 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     3961 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     4021 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     4081 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     4141 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     4201 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     4261 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     4321 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     4381 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     4441 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     4501 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     4561 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     4621 cgcctttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     4681 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     4741 tcgggtttat tacagggaca gcagagatcc agtttattta attaaggcct ccgcgccggg
     4801 ttttggcgcc tcccgcgggc gcccccctct tcacggcgag cgctgccacg tcagacgaag
     4861 ggcgcagcga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
     4921 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
     4981 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
     5041 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
     5101 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
     5161 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
     5221 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
     5281 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
     5341 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
     5401 gtggggggca tggtgggcgg caagaaccca aggtcttgag gccttcgcta atgcgggaaa
     5461 gctcttattc gggtgagatg ggctggggca ccatctgggg accctgacgt gaagtttgtc
     5521 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     5581 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     5641 ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc
     5701 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     5761 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     5821 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     5881 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     5941 tgttagacta gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacgaa
     6001 ttcgccacca tgtctagacc aaaaaagaag agaaaggtct ctagaggcga gattgctaag
     6061 tccctgaagg agattgctaa gtctctcaag gagatcgctt ggtccctgaa ggagattgct
     6121 aagtccctca agggatctag aatgatggag gccatggtga agtacctggc ggagaaagcc
     6181 ggaatcagtg aagtagaggc agcggagata gttttgaagg cggtgaaaat atctgggggt
     6241 gacgtggtga agtctataga gctggtggat ctcttcattg agatccttaa taaaggccgc
     6301 gaatggggtg gcggagggag cggtggcgga gggagcggtg gcggagggag cggtggcgga
     6361 gggagcttaa ttaagatgtc taagggggag gagcttttca caggtgtagt acctattctt
     6421 gttgaattgg atggcgatgt caatggacac aaattctccg tctcagggga gggagaggga
     6481 gacgctacct atggaaagtt gactctcaaa ttcatctgca caacgggtaa gttgccggtc
     6541 ccttggccga cactcgttac tacgttcagc tatggagtac agtgtttcgc gcgctacccc
     6601 gaccatatga aacaacacga ctttttcaaa tccgcaatgc ccgagggata cgttcaggaa
     6661 cggactattt tctttaaaga cgacggtaat tacaaaacac gcgccgaagt taagttcgaa
     6721 ggagatactt tggttaaccg aatagagctg aaaggcattg atttcaaaga ggacgggaac
     6781 atcctgggtc ataaacttga gtataatttt aattcccaca acgtgtatat aatggcagat
     6841 aaacagaaga atggcatcaa ggttaacttc aagattcggc acaatattga agacggtagt
     6901 gtacagctgg ctgaccacta tcagcagaat acgccaattg gtgatgggcc cgtcttgttg
     6961 ccagataatc actacctctc aatccagagt gcccttagta aagatccaaa cgaaaagcgg
     7021 gaccacatgg ttttgttgga gtttgtcaca gctgctggga ttacccacgg gatagacgag
     7081 ttgtataaat gactcgacgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa
     7141 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     7201 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     7261 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     7321 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     7381 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     7441 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     7501 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     7561 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     7621 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     7681 tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc
     7741 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     7801 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     7861 ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca
     7921 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     7981 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     8041 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     8101 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     8161 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     8221 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     8281 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     8341 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8401 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8461 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     8521 tgtggaaaat ctctagcagt agta
//
