LOCUS       sequence-305975-        9527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for cloning downstream of UCOE-SFFV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9527)
  AUTHORS   Landry LG, Mann SE, Anderson AM, Nakayama M
  TITLE     Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation 
            Reporter-based Detection System.
  JOURNAL   Bio Protoc. 2021 Jan 20;11(2):e3883. doi: 10.21769/BioProtoc.3883. 
            eCollection 2021 Jan 20.
  PUBMED    33732772
REFERENCE   2  (bases 1 to 9527)
  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: "Bio 
            Protoc."; date: "2021-01-20"; volume: "11(2):e3883. doi"; pages: " 
            10.21769/BioProtoc.3883. eCollection 2021 Jan 20"
COMMENT     Sequence Label: pUS
FEATURES             Location/Qualifiers
     source          1..9527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    697..814
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1099..1143)
                     /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    1328..1561
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2058..2183
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2230..2863
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    3031..3165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3055..3074)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3109..3128
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3590..3610)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3740..3805
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     promoter        complement(4939..5268)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4953..5088
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5253..5273
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(5343..5360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5514..6102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5594..5613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6273..7064,7065..7133))
                     /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     6896..6915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7134..7238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7306..7324
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7362..7384)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7484..7503
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7717..7736
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        8026..8044
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     8026..8043
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     LTR             8069..8302
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     8392..8408
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8393..8409
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8412..9000
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             8518..8529
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     8927..8947
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     promoter        complement(9075..9482)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
ORIGIN
        1 ctttttctgg agactaaata aaatctttta ttttatcggg gctgcaggac tccgcgccta
       61 cagctcaagc cacatccgaa gggggaggga gccgggagct gcgcgcgggg ccgcgcgccg
      121 cgcccgctgg gaattggggc cctaggggga gggcggaggc gccgacgacc gcggcactta
      181 ccgttcgcgg cgtggcgccc ggtggtcccc aaggggaggg aagggggagg cggggcgagg
      241 acagtgaccg gagtctcctc agcggtggct tttctgcttg gcagcctcag cggctggcgc
      301 caaaaccgga ctccgcccac ttcctcgccc ctgcggtgcg agggtgtgga atcctccaga
      361 cgctggggga gggggagttg ggagcttaaa aactagtacc cctttgggac cactttcagc
      421 agcgaactct cctgtacacc aggggtcagt tccacagacg cgggccaggg gtgggtcatt
      481 gcggcgtgaa caataatttg actagaagtt gattcgggtg tttccggaag gggccgagtc
      541 aatccgccga gttggggcac ggaaaacaaa aagggaaggc tactaagatt tttctggcgg
      601 gggttatcat tggcgtaact gcagggacca cctcccgaat tcgatatcaa gcttatcgat
      661 ccaaactgga tctctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg
      721 tttttgtaat tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc
      781 actgtacccc ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttggcg
      841 ataccgtcga gatccgttca ctaatcgaat ggatctgtct ctgtctctct ctccaccttc
      901 ttcttctatt ccttcgggcc tgtcgggtcc cctcggggtt gggaggtggg tctgaaacga
      961 taatggtgaa tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct
     1021 taaacctacc aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt
     1081 tatgttaaac caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt
     1141 cttttcttgc tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt
     1201 aatttctctg tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt
     1261 actccaacta gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca
     1321 aatccccagg agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag
     1381 ctgcttgatg ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag
     1441 caaattgttc tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt
     1501 cattgacgct gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc
     1561 tattcccact gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac
     1621 tcctaatggt tcaattttta ctactttata tttatataat tcacttctcc aattgtccct
     1681 catatctcct cctccaggtc tgaagatcag cggccggccg cttgctgtgc ggtggtctta
     1741 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     1801 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     1861 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     1921 aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     1981 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     2041 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     2101 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     2161 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     2221 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     2281 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     2341 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     2401 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     2461 tccccagtcc cgcccaggcc acgcctccct ggaaagtccc cagcggaaag tcccttgtag
     2521 caagctcgat atcagcagtt cttgaagtac tccggatgca gctctcgggc cacgtgatga
     2581 aatgctaggc ggctgtcaaa cctccactct aacacttctc tctccgggtc atccatccca
     2641 tgcaggctca cagggtgtaa caagctggtg ttctctcctt tattggcctc ttctacctta
     2701 tctggctcaa ctggtactag cttgtagcac catccaaagg tcagtggata tctgacccct
     2761 ggccctggtg tgtagttctg ctaatcaggg aagtagcctt gtgtgtggta gatccacaga
     2821 tcaaggatat cttgtcttct ttgggagtga attagccctt ccaactacta agtttgtagt
     2881 acatatttaa caaatacaat ttctttaaaa tgaaaataat tcagaggaat cacaggttta
     2941 gagtaaatga aaccacaggt aattggcagt ggtaataggg tatggggtgg gaagtttggg
     3001 atgattttgg ttagcttgag ttatccagtt gatccagaca tgataagata cattgatgag
     3061 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3121 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3181 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3241 ctctacaaat gtggtatggc tgattatgat ctctagtcaa ggcactatac atcaaatatt
     3301 ccttattaac ccctttacaa attaaaaagc taaaggtaca caatttttga gcatagttat
     3361 taatagcaga cactctatgc ctgtgtggag taagaaaaaa cagtatgtta tgattataac
     3421 tgttatgcct acttataaag gttacagaat atttttccat aattttcttg tatagcagtg
     3481 cagctttttc ctttgtggtg taaatagcaa agcaagcaag agttctatta ctaaacacag
     3541 catgactcaa aaaacttagc aattctgaag gaaagtcctt ggggtcttct acctttctct
     3601 tcttttttgg aggagtagaa tgttgagagt cagcagtagc ctcatcatca ctagatggca
     3661 tttcttctga gcaaaacagg ttttcctcat taaaggcatt ccaccactgc tcccattcat
     3721 cagttccata ggttggaatc taaaatacac aaacaattag aatcagtagt ttaacacatt
     3781 atacacttaa aaattttata tttaccttag agctttaaat ctctgtaggt agtttgtcca
     3841 attatgtcac accacagaag taaggttcct tcacaaagat ccggggccca ctcataaatc
     3901 cagttgccgc cacggtagcc aatcaccgta tcgtataaat catcgtcggt acgttcggca
     3961 tcgctcatca caatacgtgc ctggacgtcg aggatttcgc gtgggtcaat gccgcgccag
     4021 atccacatca gacggttaat catgcgatac cagtgaggga tggttttacc atcaagggcc
     4081 gactgcacag gcggttgtgc gccgtgatta aagcggcgga ctagcgtcga ggtttcagga
     4141 tgtttaaagc ggggtttgaa cagggtttcg ctcaggtttg cctgtgtcat ggatgcagcc
     4201 tccagaatac ttactggaaa ctattgtaac ccgcctgaag ttaaaaagaa caacgcccgg
     4261 cagtgccagg cgttgaaaag attagcgacc ggagattggc gggacgaata cgacgcccat
     4321 atcccacggc tgttcaatcc aggtatcttg cgggatatca acaacatagt catcaaccag
     4381 cggacgacca gccggttttg cgaagatggt gacaaagtgc gcttttggat acatttcacg
     4441 aatcgcaacc gcagtaccac cggtatccac caggtcatca ataacgatga agccttcgcc
     4501 atcgccttct gcgcgtttca gcactttaag ctcgcgctgg ttgtcgtgat cgtagctgga
     4561 aatacaaacg gtatcgacat gacgaatacc cagttcacgc gccagtaacg cacccggtac
     4621 agaccgccac ggcttacggc aataatgcct ttccattgtt cagaaggcat cagtcggctt
     4681 gcgagtttac gtgcatggat ctgcaacatg tcccaggtga cgatgtattt ttcgctcatg
     4741 tgaagtgtcc cagcctgttt atctacggct taaaaagtgt tcgaggggaa aataggttgc
     4801 gcgagattat agagatctgg cgcactaaaa accagtattt cacatgagtc cgcgtctttt
     4861 tacgcactgc ctctccctga cgcgggataa agtggtattc tcaaacatat ctcgcaagcc
     4921 tgtcttgtgt ccaagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     4981 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     5041 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     5101 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5161 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     5221 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     5281 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5341 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5401 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5461 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5521 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5581 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5641 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5701 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5761 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5821 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5881 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5941 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6001 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6061 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6121 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6181 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6241 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6301 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6361 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6421 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6481 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6541 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6601 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6661 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6721 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6781 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6841 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6901 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6961 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7021 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7081 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7141 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7201 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7261 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7321 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     7381 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     7441 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     7501 gtactgagag tgcaccattc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     7561 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     7621 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     7681 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     7741 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg
     7801 gctagcgatg accctgctga ttggttcgct gaccatttcc gggtgcggga cggcgttacc
     7861 agaaactcag aaggttcgtc caaccaaacc gactctgacg gcagtttacg agagagatga
     7921 tagggtctgc ttcagtaagc cagatgctac acaattaggc ttgtacatat tgtcgttaga
     7981 acgcggctac aattaataca taaccttatg tatcatacac atacgattta ggtgacacta
     8041 tagaatacac ggaattaatt ctagatgctg ctagagattt tccacactga ctaaaagggt
     8101 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact
     8161 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag
     8221 gctcagatct ggtctaacca gagagaccca gtacaagcaa aaagcagatc ttgtcttcgt
     8281 tgggagtgaa ttagcccttc cagtcccccc ttttctttta aaaagtggct aagatctaca
     8341 gctgccttgt aagtcattgg tcttaaaggt accgagctcg aattaattcc ctcgaggtcg
     8401 acggtatcga tgcggggagg cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg
     8461 aagacgcgga agaggccgca gagccggcag caggccgcgg gaaggaaggt ccgctggatt
     8521 gagggccgaa gggacgtagc agaaggacgt cccgcgcaga atccaggtgg caacacaggc
     8581 gagcagccaa ggaaaggacg atgatttccc cgacaacacc acggaattgt cagtgcccaa
     8641 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaat
     8701 agggaggggg aaagcgaaag tcccggaaag gagctgacag gtggtggcaa tgccccaacc
     8761 agtgggggtt gcgtcagcaa acacagtgca caccacgcca cgttgcctga caacgggcca
     8821 caactcctca taaagagaca gcaaccagga tttatacaag gaggagaaaa tgaaagccat
     8881 acgggaagca atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg
     8941 agcaacatag ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt
     9001 atcgataagc ttgatatcaa gcttgcatgc ctgcagggta gacgcgtgag aattctcgag
     9061 agctccagat cccccccggg cgactcagtc tgtcggagga ctggcgcgcc gagtgagggg
     9121 ttgtgagctc ttttatagag ctcgggaagc agaagcgcgc gaacagaagc gagaagcagg
     9181 ctgattggtt aattcaaata aggcgcaggg tcatttcagg tccttggggg agcctggaaa
     9241 catctgatgg gtcttaagaa actgctgagg gttgggccat atctggggac catctgttct
     9301 tggcctcggg ccggggccga aactgcggtg accatctgtt cttggccccg ggccggggcc
     9361 gaaactgctc accgcagata tcctgtttgg cccaacgtta gctattttca tgtacccgcc
     9421 cttgatctga acttctctat tcttggtttg gtatttttcc atgccttgca aaatggcgtt
     9481 actgcagcta gcttgccaaa cctacaggtg gggtctttca ttccccc
//
