LOCUS       sequence-402484-        9294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian lentiviral vector for the expression of the 
            Sapphire-tagged nuclear localizing 40nm nucGEMs under the UBC 
            promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9294)
  AUTHORS   Shu T et al.
  TITLE     nucGEMs probe the biophysical properties of the nucleoplasm
  JOURNAL   bioRxiv 2022
REFERENCE   2  (bases 1 to 9294)
  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"; journalName: "bioRxiv 
            2022"
COMMENT     Sequence Label: pUBC-SV40NLS-Pfv-Sapphire
FEATURES             Location/Qualifiers
     source          1..9294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             24..657
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    704..829
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1326..1559
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1744..1788
                     /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    2051..2166
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2212..3423
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3260..3279
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3445..3465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5286..5874
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5339..5359)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5757..5768)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6081..6714
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6842..6858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6842..6858)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6855..6877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6866..6882
                     /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(6890..6896,6897..6914,6915..6920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6935..6956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7073..7090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7244..7832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7324..7343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8003..8794,8795..8863))
                     /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     8626..8645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8864..8968)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9016..9150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9053..9072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9107..9126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 atatgtacta caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat
       61 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
      121 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
      181 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
      241 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
      301 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
      361 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
      421 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt
      481 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
      541 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
      601 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
      661 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
      721 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
      781 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
      841 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
      901 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
      961 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     1021 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     1081 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     1141 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga
     1201 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1261 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1321 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1381 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1441 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1501 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1561 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1621 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1681 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1741 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1801 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1861 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1921 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1981 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     2041 cggtatcgcc tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2101 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2161 aaattttcgg gtttattaca gggacagcag agatccagtt tatttaatta aggcctccgc
     2221 gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag
     2281 acgaagggcg cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct
     2341 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact
     2401 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt
     2461 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata
     2521 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct
     2581 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc
     2641 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg
     2701 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat
     2761 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga
     2821 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc
     2881 gggaaagctc ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag
     2941 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     3001 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     3061 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     3121 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     3181 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     3241 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     3301 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     3361 tttcagtgtt agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     3421 gacgaattcg ccaccatgtc tagaccaaaa aagaagagaa aggtctctag actctctatc
     3481 aacccgacat tgataaaccg ggacaaaccg tatactaaag aagaacttat ggaaatcctg
     3541 cggctggcaa tcattgctga gttggacgcg atcaacctct atgaacagat ggcacggtat
     3601 tctgaagatg agaacgtaag gaagattctc cttgacgtcg ctcgggagga gaaggcgcac
     3661 gtgggtgagt ttatggcact ccttcttaac cttgaccctg agcaagtgac tgagttgaaa
     3721 ggcggcttcg aagaggtaaa agaactgaca ggcattgaag cccatattaa cgataataaa
     3781 aaagaagagt ccaacgtcga atattttgaa aaactccgct ccgctctgtt ggatggagta
     3841 aataagggga ggagtctcct taagcatctg ccagttacac gcatagaagg gcagtccttt
     3901 cgagtagaca ttattaaatt tgaggatggg gtgcgcgtcg ttaaacagga atacaagccc
     3961 attccacttc tcaaaaagaa attttacgta ggaattagag aactgaatga tggtacgtac
     4021 gacgtttcta ttgccacgaa ggcaggtgaa ttgttggtta aggacgagga aagccttgtg
     4081 ataagagaaa tcctgtcaac tgaagggata aagaaaatga agttgagtag ctgggacaat
     4141 cccgaggagg cgctgaatga tcttatgaat gcccttcagg aagcctctaa tgcttccgct
     4201 ggacctttcg gtctgataat taatccaaag agatacgcga aacttcttaa aatctatgag
     4261 aaatcaggaa aaatgttggt agaagtgctg aaagaaattt ttcggggtgg catcatagtc
     4321 acactgaaca tagatgaaaa caaggttatt atttttgcaa atactccagc ggtgcttgac
     4381 gtagttgttg ggcaggacgt aacgctccag gaattgggac ctgaggggga cgacgtggcc
     4441 tttttggtgt cagaagccat cggcatccga ataaagaacc ccgaagcgat tgttgtcctc
     4501 gaatggtcag ggtcaggttc tggctcaggt agcgggagtg ggcggccgct gatgtctaag
     4561 ggggaggagc ttttcacagg tgtagtacct attcttgttg aattggatgg cgatgtcaat
     4621 ggacacaaat tctccgtctc aggggaggga gagggagacg ctacctatgg aaagttgact
     4681 ctcaaattca tctgcacaac gggtaagttg ccggtccctt ggccgacact cgttactacg
     4741 ttcagctatg gagtacagtg tttcgcgcgc taccccgacc atatgaaaca acacgacttt
     4801 ttcaaatccg caatgcccga gggatacgtt caggaacgga ctattttctt taaagacgac
     4861 ggtaattaca aaacacgcgc cgaagttaag ttcgaaggag atactttggt taaccgaata
     4921 gagctgaaag gcattgattt caaagaggac gggaacatcc tgggtcataa acttgagtat
     4981 aattttaatt cccacaacgt gtatataatg gcagataaac agaagaatgg catcaaggtt
     5041 aacttcaaga ttcggcacaa tattgaagac ggtagtgtac agctggctga ccactatcag
     5101 cagaatacgc caattggtga tgggcccgtc ttgttgccag ataatcacta cctctcaatc
     5161 cagagtgccc ttagtaaaga tccaaacgaa aagcgggacc acatggtttt gttggagttt
     5221 gtcacagctg ctgggattac ccacgggata gacgagttgt ataaatgact cgacgcgtct
     5281 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5341 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5401 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5461 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5521 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5581 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5641 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     5701 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5761 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5821 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     5881 ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
     5941 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
     6001 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6061 tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct
     6121 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     6181 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     6241 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     6301 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     6361 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     6421 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     6481 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     6541 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     6601 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6661 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6721 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6781 agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg gcgtaatcat
     6841 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6901 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6961 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     7021 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7081 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7141 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7201 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7261 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7321 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7381 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7441 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7501 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7561 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7621 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7681 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7741 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7801 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7861 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7921 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7981 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8041 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8101 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8161 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8221 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8281 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8341 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8401 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8461 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8521 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8581 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8641 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8701 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8761 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8821 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8881 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8941 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     9001 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     9061 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9121 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
     9181 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
     9241 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaa
//
