LOCUS       sequence-261172-        8502 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  nucleus marker, lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8502)
  TITLE     Okada Lab plasmids
REFERENCE   2  (bases 1 to 8502)
  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"
COMMENT     Sequence Label: pLBS.CAG/NLS-mScarlet
FEATURES             Location/Qualifiers
     source          1..8502
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      complement(22..477)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(159..180)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     371..390
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     604..626
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     618..635
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     619..635
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     642..661
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        642..660
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        680..906
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             907..1087
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1134..1259
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1752..1985
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2170..2214
                     /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    2481..2597
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2676..2979
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2981..3256
                     /label=chicken beta-actin promoter
     intron          3257..4273
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4197..4219
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4281..4300
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4349..4369
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4373..4393
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4397..4417
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4436..5131
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(4569..4587)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4837..4856
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5186..5774
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5239..5259)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5657..5668)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5865..6098
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6170..6251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6207..6226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6261..6280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(6329..6347)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6368..6384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6368..6384)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6381..6403)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6392..6408
                     /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(6416..6422,6423..6440,6441..6446))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6461..6482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6599..6616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6770..7358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6850..6869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7529..8320,8321..8389))
                     /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     8152..8171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8390..8494)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
       61 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
      121 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
      181 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
      241 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
      301 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
      361 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
      421 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
      481 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      541 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      601 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
      661 gcgaattggg taccacgcgt gtagtcttat gcaatactct tgtagtcttg caacatggta
      721 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
      781 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
      841 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
      901 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      961 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     1021 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     1081 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     1141 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     1201 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     1261 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     1321 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1381 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1441 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1501 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1561 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     1621 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     1681 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     1741 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     1801 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     1861 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     1921 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     1981 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     2041 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     2101 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     2161 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     2221 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     2281 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     2341 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     2401 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     2461 tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     2521 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2581 acaaaattca aaattttggg cccttccaac cttccaaccg gtacctagta atcaattacg
     2641 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2701 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2761 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2821 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2881 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2941 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
     3001 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
     3061 taattatttt gtgcagcgat gggggcgggg gggggggggg cgcgcgccrg gcggggcggg
     3121 gcggggcggg gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
     3181 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
     3241 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
     3301 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
     3361 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
     3421 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
     3481 ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc
     3541 cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga
     3601 ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg
     3661 ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct
     3721 gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg
     3781 gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg
     3841 ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg
     3901 cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta
     3961 atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg
     4021 gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa
     4081 ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca
     4141 gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
     4201 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
     4261 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
     4321 aggattcctc gatgccacca tgctggtacc aaaaaagaag agaaaggttg taccaaaaaa
     4381 gaagagaaag gttgtaccaa aaaagaagag aaaggttttc gaagggccgg atccaatggt
     4441 gagcaagggc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc
     4501 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     4561 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct
     4621 gtcccctcag ttcatgtacg gctccagggc cttcaccaag caccccgccg acatccccga
     4681 ctactataag cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     4741 cggcggcgcc gtgaccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa
     4801 ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat
     4861 gggctgggaa gcgtccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat
     4921 taagatggcc ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta
     4981 caaggccaag aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca agttggacat
     5041 cacctcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca
     5101 ctccaccggc ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttg
     5161 agaattcgac tagtctcgac tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5221 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5281 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5341 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5401 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5461 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5521 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5581 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5641 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5701 ggctctgcgg cctcttccgc atcttcgcct tcgccctcag acgagtcgga tctccctttg
     5761 ggccgcctcc ccgcctggaa ttaattcgag ctcggtacct ttaagaccaa tgacttacaa
     5821 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     5881 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     5941 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6001 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6061 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     6121 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     6181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatcta gagcggccgc
     6301 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     6361 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6421 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6601 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6661 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6721 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6781 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6841 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6901 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6961 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7021 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7081 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7141 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7201 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7261 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7381 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7441 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7501 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7561 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7621 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7681 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7741 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7801 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7861 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7921 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7981 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8041 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8101 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8161 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8221 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8281 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8341 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8461 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tc
//
