LOCUS       sequence-444087-        9617 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral expression of human CD28 (Y209F) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9617)
  AUTHORS   Zhao Y, Caron C, Chan YY, Lee CK, Xu X, Zhang J, Masubuchi T, Wu C, 
            Bui JD, Hui E
  TITLE     cis-B7:CD28 interactions at invaginated synaptic membranes provide 
            CD28 co-stimulation and promote CD8(+) T cell function and 
            anti-tumor immunity.
  JOURNAL   Immunity. 2023 Jun 13;56(6):1187-1203.e12. doi: 
            10.1016/j.immuni.2023.04.005. Epub 2023 May 8.
  PUBMED    37160118
REFERENCE   2  (bases 1 to 9617)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.immuni.2023.04.005"; journalName: "Immunity"; date: 
            "2023-06-13- 13"; volume: "56"; issue: "6"; pages: "1187-1203.e12"
COMMENT     Sequence Label: pHR-CD28(Y209F)
FEATURES             Location/Qualifiers
     source          1..9617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..17)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2..18)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             108..341
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(366..384)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(367..384)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(674..693)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(907..926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1026..1048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1086..1104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1172..1276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1277..1345,1346..2137)
                     /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     complement(1495..1514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2308..2896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2797..2816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3050..3067
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3137..3157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3142..3471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3322..3457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4605..4670
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4800..4820
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5245..5379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5282..5301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5336..5355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5547..6180
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6227..6352
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6849..7082
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7267..7311
                     /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    7596..7713
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7866..8273
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    9027..9615
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9080..9100)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9498..9509)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 cgataccgtc gacctcgagg gaattaattc gagctcggta cctttaagac caatgactta
       61 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
      121 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
      181 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
      241 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
      301 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatctaga attaattccg
      361 tgtattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg
      421 tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg tagcatctgg cttactgaag
      481 cagaccctat catctctctc gtaaactgcc gtcagagtcg gtttggttgg acgaaccttc
      541 tgagtttctg gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat cagcagggtc
      601 atcgctagcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
      661 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
      721 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
      781 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
      841 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg aatggtgcac
      901 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
      961 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1021 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     1081 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1141 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1201 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1261 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1321 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1381 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1441 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1501 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1561 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1621 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1681 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1741 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1801 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1861 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1921 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1981 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2041 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2101 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2161 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2221 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2281 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2341 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2401 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2461 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2521 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2581 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2641 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2701 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2761 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2821 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2881 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2941 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3001 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3061 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3121 tcattaatgc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgga
     3481 cacaagacag gcttgcgaga tatgtttgag aataccactt tatcccgcgt cagggagagg
     3541 cagtgcgtaa aaagacgcgg actcatgtga aatactggtt tttagtgcgc cagatctcta
     3601 taatctcgcg caacctattt tcccctcgaa cactttttaa gccgtagata aacaggctgg
     3661 gacacttcac atgagcgaaa aatacatcgt cacctgggac atgttgcaga tccatgcacg
     3721 taaactcgca agccgactga tgccttctga acaatggaaa ggcattattg ccgtaagccg
     3781 tggcggtctg taccgggtgc gttactggcg cgtgaactgg gtattcgtca tgtcgatacc
     3841 gtttgtattt ccagctacga tcacgacaac cagcgcgagc ttaaagtgct gaaacgcgca
     3901 gaaggcgatg gcgaaggctt catcgttatt gatgacctgg tggataccgg tggtactgcg
     3961 gttgcgattc gtgaaatgta tccaaaagcg cactttgtca ccatcttcgc aaaaccggct
     4021 ggtcgtccgc tggttgatga ctatgttgtt gatatcccgc aagatacctg gattgaacag
     4081 ccgtgggata tgggcgtcgt attcgtcccg ccaatctccg gtcgctaatc ttttcaacgc
     4141 ctggcactgc cgggcgttgt tctttttaac ttcaggcggg ttacaatagt ttccagtaag
     4201 tattctggag gctgcatcca tgacacaggc aaacctgagc gaaaccctgt tcaaaccccg
     4261 ctttaaacat cctgaaacct cgacgctagt ccgccgcttt aatcacggcg cacaaccgcc
     4321 tgtgcagtcg gcccttgatg gtaaaaccat ccctcactgg tatcgcatga ttaaccgtct
     4381 gatgtggatc tggcgcggca ttgacccacg cgaaatcctc gacgtccagg cacgtattgt
     4441 gatgagcgat gccgaacgta ccgacgatga tttatacgat acggtgattg gctaccgtgg
     4501 cggcaactgg atttatgagt gggccccgga tctttgtgaa ggaaccttac ttctgtggtg
     4561 tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt
     4621 taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct
     4681 atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct
     4741 cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc
     4801 caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt
     4861 tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac accacaaagg
     4921 aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc tttataagta
     4981 ggcataacag ttataatcat aacatactgt tttttcttac tccacacagg catagagtgt
     5041 ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt tgtaaagggg
     5101 ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag ccataccaca
     5161 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5221 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5341 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     5401 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     5461 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     5521 taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa gaagacaaga
     5581 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
     5641 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     5701 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
     5761 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
     5821 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
     5881 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
     5941 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
     6001 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6061 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6121 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6181 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     6241 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     6301 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     6361 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     6421 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     6481 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     6541 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     6601 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     6661 aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga
     6721 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     6781 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     6841 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     6901 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     6961 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     7021 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     7081 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     7141 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     7201 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     7261 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     7321 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     7381 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     7441 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     7501 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     7561 cgacggtatc gccaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     7621 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     7681 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     7741 ccagtttgga tcgataagct tgatatcgaa ttcctgcagc cccgataaaa taaaagattt
     7801 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     7861 ctgcagtaac gccattttgc aaggcatgga aaaataccaa accaagaata gagaagttca
     7921 gatcaagggc gggtacatga aaatagctaa cgttgggcca aacaggatat ctgcggtgag
     7981 cagtttcggc cccggcccgg ggccaagaac agatggtcac cgcagtttcg gccccggccc
     8041 gaggccaaga acagatggtc cccagatatg gcccaaccct cagcagtttc ttaagaccca
     8101 tcagatgttt ccaggctccc ccaaggacct gaaatgaccc tgcgccttat ttgaattaac
     8161 caatcagcct gcttctcgct tctgttcgcg cgcttctgct tcccgagctc tataaaagag
     8221 ctcacaaccc ctcactcggc gcgccagtcc tccgacagac tgagtcgccc gggggggatc
     8281 tggagctctc gagaattctc acgatgctca ggctgctctt ggctctcaac ttattccctt
     8341 caattcaagt aacaggaaac aagattttgg tgaagcagtc gcccatgctt gtagcgtacg
     8401 acaatgcggt caaccttagc tgcaagtatt cctacaatct cttctcaagg gagttccggg
     8461 catcccttca caaaggactg gatagtgctg tggaagtctg tgttgtatat gggaattact
     8521 cccagcagct tcaggtttac tcaaaaacgg ggttcaactg tgatgggaaa ttgggcaatg
     8581 aatcagtgac attctacctc cagaatttgt atgttaacca aacagatatt tacttctgca
     8641 aaattgaagt tatgtatcct cctccttacc tagacaatga gaagagcaat ggaaccatta
     8701 tccatgtgaa agggaaacac ctttgtccaa gtcccctatt tcccggacct tctaagccct
     8761 tttgggtgct ggtggtggtt ggtggagtcc tggcttgcta tagcttgcta gtaacagtgg
     8821 cctttattat tttctgggtg aggagtaaga ggagcaggct cctgcacagt gactacatga
     8881 acatgactcc ccgccgcccc gggcccaccc gcaagcatta ccagcccttc gccccaccac
     8941 gcgacttcgc agcctatcgc tcctaaagcg gccgcgactc tagagtcgac ctgcaggcat
     9001 gcaagcttga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     9061 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     9121 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     9181 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     9241 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     9301 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     9361 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     9421 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     9481 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     9541 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     9601 gggccgcctc cccgcat
//
