LOCUS       sequence-327887-        7951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CMV UL83 (363-373) fused to IL-2 signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7951)
  AUTHORS   Lee MN, Meyerson M
  TITLE     Antigen identification for HLA class I- and HLA class II-restricted 
            T cell receptors using cytokine-capturing antigen-presenting cells.
  JOURNAL   Sci Immunol. 2021 Jan 22;6(55). pii: 6/55/eabf4001. doi: 
            10.1126/sciimmunol.abf4001.
  PUBMED    33483338
REFERENCE   2  (bases 1 to 7951)
  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: "Sci 
            Immunol."; date: "2021-01-22"; volume: "6(55). pii: 6/55/eabf4001. 
            doi"; pages: " 10.1126/sciimmunol.abf4001"
COMMENT     Sequence Label: YSEHPTFTSQY_epitope
FEATURES             Location/Qualifiers
     source          1..7951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     sig_peptide     612..671
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     misc_feature    762..1350
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(815..835)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1233..1244)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1422..1655
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1733..1854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1770..1789)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1824..1843
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1894..2029
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2049..2068)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2050..2068)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2078..2095)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2078..2094)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2087..2109)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2236..2691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2323..2342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2533..2554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2717..2821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2822..2890,2891..3682)
                     /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(3040..3059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3853..4441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4342..4361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4595..4612
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4729..4750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4765..4770,4771..4788,4789..4795)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4803..4819
                     /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)."
     primer_bind     4808..4830
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4827..4843
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4827..4843
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4862..4882
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4864..4882
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4910..5136
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5137..5317
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5364..5489
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5982..6215
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6400..6444
                     /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"
     promoter        6619..7119
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7038..7056
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7141..7740
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7141..7160)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7637..7657
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7782..7899
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccggggtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactgtcggg atcaacaagt
      601 ttgccgccac catgtacagg atgcaactcc tgtcttgcat tgcactaagt cttgcacttg
      661 tcacgaattc atatagcgag caccctacct tcaccagcca gtattgaatt caaagtggtt
      721 tagtaatgag ctagcgctaa ccggtggcgc gttaagtcga caatcaacct ctggattaca
      781 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
      841 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
      901 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
      961 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     1021 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     1081 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     1141 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     1201 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     1261 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     1321 gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc
     1381 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     1441 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     1501 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     1561 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     1621 cagacccttt tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt catgtcatct
     1681 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     1741 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1801 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1861 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     1921 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     1981 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac
     2041 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     2101 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2161 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2221 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2281 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2341 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2401 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2461 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2521 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2581 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2641 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     2701 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2761 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2821 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2881 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2941 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3001 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3061 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3121 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3181 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3241 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3301 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3361 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3421 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3481 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3541 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3601 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3661 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3721 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3781 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3841 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3901 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3961 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4021 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4081 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4141 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4201 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4261 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4321 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4381 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4441 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4501 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4561 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4621 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     4681 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     4741 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     4801 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
     4861 cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat
     4921 gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga
     4981 gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta
     5041 ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga
     5101 gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc
     5161 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5221 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5281 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     5341 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     5401 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     5461 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     5521 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     5581 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     5641 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     5701 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     5761 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     5821 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     5881 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     5941 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     6001 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     6061 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     6121 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     6181 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     6241 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     6301 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     6361 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     6421 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     6481 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     6541 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     6601 ccaaccccga ggggaccctt gcgccttttc caaggcagcc ctgggtttgc gcagggacgc
     6661 ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggtc tcgcacattc
     6721 ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg ccccccggcg
     6781 acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc gtgccggacg
     6841 tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg ccagggagca
     6901 atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc agggcgcgcc
     6961 gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc
     7021 ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc
     7081 ggctccctcg ttgaccgaat caccgacctc tctccccagg gggtaccacc ggagcttacc
     7141 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     7201 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     7261 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     7321 atcggcaagg tgtgggtcgc ggacgacggc gccgccgtgg cggtctggac cacgccggag
     7381 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     7441 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     7501 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     7561 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     7621 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     7681 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     7741 gaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     7801 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     7861 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     7921 agagatccac tttggcgccg gctcgagggg g
//
