LOCUS       sequence-444955-        6096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of mouse CD28 (Y189F) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  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 6096)
  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"; 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: pQCX-CD28 Y189F
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     463..483
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1627..1924
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     1947..1966
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1947..1965
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           1980..2022
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     LTR             2802..3227
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(3330..3352)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3493..3822
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3673..3808
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3844..3863
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3944..3961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4115..4703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4195..4214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4874..5665,5666..5734))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5497..5516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5735..5839)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5907..5925
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(6025..6096,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tcccaaccag cacagtggtc gaccctgtgg aatgtgtgtc
     1621 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     1681 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     1741 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     1801 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     1861 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     1921 ttggaggatc ttaaggctag agtacttaat acgactcact ataggggcaa acaaaattgc
     1981 ttgttctttt tgcagaagct cagaataaac gctcaacttt ggcagatacg cgtgccacca
     2041 tgacactcag gctgctgttc ttggctctca acttcttctc agttcaagta acagaaaaca
     2101 agattttggt aaagcagtcg cccctgcttg tggtagatag caacgaggtc agcctcagct
     2161 gcaggtattc ctacaacctt ctcgcaaagg aattccgggc atccctgtac aagggcgtga
     2221 acagcgacgt ggaagtctgt gtcgggaatg ggaattttac ctatcagccc cagtttcgct
     2281 cgaatgccga gttcaactgc gacggggatt tcgacaacga aacagtgacg ttccgtctct
     2341 ggaatctgca cgtcaatcac acagatattt acttctgcaa aattgagttc atgtaccctc
     2401 cgccttacct agacaacgag aggagcaatg gaactattat tcacataaaa gagaaacatc
     2461 tttgtcatac tcagtcatct cctaagctgt tttgggcact ggtcgtggtt gctggagtcc
     2521 tgttttgtta tggcttgcta gtgacagtgg ctctttgtgt tatctggaca aatagtagaa
     2581 ggaacagact ccttcaaagt gacttcatga acatgactcc ccggaggcct gggctcactc
     2641 gaaagcctta ccagccctac gcccctgcca gagactttgc agcgtaccgc ccctaagcgg
     2701 ccgcatctta attaagaatt atcaagctta tcgctgaagc ctatagagta cgagccatag
     2761 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     2821 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     2881 agaatagaga agttcagatc aaggtcagga acagatggaa cagggtcgac cctagagaac
     2941 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     3001 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     3061 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     3121 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     3181 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc
     3241 gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc
     3301 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3361 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3421 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtagatc
     3481 cggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3541 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3721 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac tggcttaact
     3841 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     3901 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     3961 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4021 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4081 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4141 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4201 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4261 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4321 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4381 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4441 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4501 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4561 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4621 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4681 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4741 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4801 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4861 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4921 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4981 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5041 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5101 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5161 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     5221 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5281 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5341 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5401 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5461 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     5521 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5581 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5641 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5701 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5761 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5821 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5881 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     5941 aattagcttg gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct
     6001 catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
     6061 ttacggggtc attagttcat agcccatata tggagt
//
