LOCUS       sequence-402479-        9267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian lentiviral vector for the expression of the 
            Sapphire-tagged 40nm GEM under the UBC promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9267)
  AUTHORS   Shu T et al.
  TITLE     nucGEMs probe the biophysical properties of the nucleoplasm
  JOURNAL   bioRxiv 2022
REFERENCE   2  (bases 1 to 9267)
  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-Pfv-Sapphire
FEATURES             Location/Qualifiers
     source          1..9267
                     /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    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2189..3400
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3237..3256
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     misc_feature    5236..5824
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5289..5309)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5707..5718)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6031..6664
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6792..6808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6792..6808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6805..6827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6816..6832
                     /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(6840..6846,6847..6864,6865..6870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6885..6906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7023..7040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7194..7782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7274..7293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7953..8744,8745..8813))
                     /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     8576..8595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8814..8918)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8966..9100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9003..9022)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9057..9076
                     /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 tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat ttaattaagg cctccgcgcc gggttttggc gcctcccgcg
     2221 ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct
     2281 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     2341 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     2401 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     2461 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     2521 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     2581 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     2641 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     2701 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     2761 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     2821 cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta ttcgggtgag
     2881 atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     2941 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     3001 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     3061 agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     3121 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     3181 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     3241 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     3301 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt
     3361 gtccgctaaa ttctggccgt ttttggcttt tttgttagac gaattcgcca ccatgtctag
     3421 actctctatc aacccgacat tgataaaccg ggacaaaccg tatactaaag aagaacttat
     3481 ggaaatcctg cggctggcaa tcattgctga gttggacgcg atcaacctct atgaacagat
     3541 ggcacggtat tctgaagatg agaacgtaag gaagattctc cttgacgtcg ctcgggagga
     3601 gaaggcgcac gtgggtgagt ttatggcact ccttcttaac cttgaccctg agcaagtgac
     3661 tgagttgaaa ggcggcttcg aagaggtaaa agaactgaca ggcattgaag cccatattaa
     3721 cgataataaa aaagaagagt ccaacgtcga atattttgaa aaactccgct ccgctctgtt
     3781 ggatggagta aataagggga ggagtctcct taagcatctg ccagttacac gcatagaagg
     3841 gcagtccttt cgagtagaca ttattaaatt tgaggatggg gtgcgcgtcg ttaaacagga
     3901 atacaagccc attccacttc tcaaaaagaa attttacgta ggaattagag aactgaatga
     3961 tggtacgtac gacgtttcta ttgccacgaa ggcaggtgaa ttgttggtta aggacgagga
     4021 aagccttgtg ataagagaaa tcctgtcaac tgaagggata aagaaaatga agttgagtag
     4081 ctgggacaat cccgaggagg cgctgaatga tcttatgaat gcccttcagg aagcctctaa
     4141 tgcttccgct ggacctttcg gtctgataat taatccaaag agatacgcga aacttcttaa
     4201 aatctatgag aaatcaggaa aaatgttggt agaagtgctg aaagaaattt ttcggggtgg
     4261 catcatagtc acactgaaca tagatgaaaa caaggttatt atttttgcaa atactccagc
     4321 ggtgcttgac gtagttgttg ggcaggacgt aacgctccag gaattgggac ctgaggggga
     4381 cgacgtggcc tttttggtgt cagaagccat cggcatccga ataaagaacc ccgaagcgat
     4441 tgttgtcctc gaatggtcag ggtcaggttc tggctcaggt agcgggagtg ggcggccgct
     4501 gatgtctaag ggggaggagc ttttcacagg tgtagtacct attcttgttg aattggatgg
     4561 cgatgtcaat ggacacaaat tctccgtctc aggggaggga gagggagacg ctacctatgg
     4621 aaagttgact ctcaaattca tctgcacaac gggtaagttg ccggtccctt ggccgacact
     4681 cgttactacg ttcagctatg gagtacagtg tttcgcgcgc taccccgacc atatgaaaca
     4741 acacgacttt ttcaaatccg caatgcccga gggatacgtt caggaacgga ctattttctt
     4801 taaagacgac ggtaattaca aaacacgcgc cgaagttaag ttcgaaggag atactttggt
     4861 taaccgaata gagctgaaag gcattgattt caaagaggac gggaacatcc tgggtcataa
     4921 acttgagtat aattttaatt cccacaacgt gtatataatg gcagataaac agaagaatgg
     4981 catcaaggtt aacttcaaga ttcggcacaa tattgaagac ggtagtgtac agctggctga
     5041 ccactatcag cagaatacgc caattggtga tgggcccgtc ttgttgccag ataatcacta
     5101 cctctcaatc cagagtgccc ttagtaaaga tccaaacgaa aagcgggacc acatggtttt
     5161 gttggagttt gtcacagctg ctgggattac ccacgggata gacgagttgt ataaatgact
     5221 cgacgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     5281 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     5341 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     5401 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5461 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5521 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5581 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     5641 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5701 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5761 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5821 ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca
     5881 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     5941 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6001 ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac
     6061 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
     6121 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
     6181 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
     6241 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
     6301 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
     6361 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
     6421 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
     6481 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6541 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6601 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6661 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     6721 tcagagagtg agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg
     6781 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6841 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6901 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6961 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7021 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7081 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7141 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7201 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7261 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7321 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7381 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7441 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7501 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7561 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7621 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7681 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7741 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7801 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7861 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7921 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7981 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8041 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8101 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8161 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8221 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8281 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8341 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8401 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8461 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8521 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8581 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8641 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8701 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8761 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8821 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8881 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8941 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
     9001 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     9061 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
     9121 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
     9181 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
     9241 ttaaatatgt actacaaact tagtagt
//
