LOCUS       sequence-370614-        7944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CMV UL83 (116-123) fused to IL-2 signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7944)
  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 7944)
  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: LPLKMLNI_epitope
FEATURES             Location/Qualifiers
     source          1..7944
                     /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    755..1343
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(808..828)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1226..1237)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1415..1648
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1726..1847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1763..1782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1817..1836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1887..2022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2042..2061)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2043..2061)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2071..2088)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2071..2087)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2080..2102)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2229..2684
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2316..2335)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2526..2547
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2710..2814
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2815..2883,2884..3675)
                     /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(3033..3052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3846..4434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4335..4354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4588..4605
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4722..4743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4758..4763,4764..4781,4782..4788)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4796..4812
                     /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     4801..4823
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4820..4836
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4820..4836
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4855..4875
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4857..4875
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4903..5129
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5130..5310
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5357..5482
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5975..6208
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6393..6437
                     /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        6612..7112
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7031..7049
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7134..7733
                     /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(7134..7153)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7630..7650
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7775..7892
                     /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 tgctctttcc ctcgccttag
      661 taacgaattc actgcctctg aagatgctga acatctgaat tcaaagtagg tttgttaagt
      721 aagctagcgc taaccggtgg cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
      781 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
      841 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
      901 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
      961 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1021 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     1081 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     1141 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     1201 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     1261 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     1321 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     1381 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     1441 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     1501 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     1561 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     1621 ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt
     1681 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     1741 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1801 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     1861 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1921 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     1981 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     2041 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     2101 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     2161 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     2221 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2281 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2341 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2401 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2461 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2521 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2581 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2641 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     2701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2761 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2821 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2881 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2941 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3001 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3061 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3121 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3181 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3241 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3301 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3361 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3421 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3481 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3541 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3601 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3661 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3721 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3781 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3841 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3901 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3961 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4021 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4081 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4141 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4201 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4261 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4321 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4381 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4441 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4501 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4561 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     4621 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     4681 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     4741 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     4801 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     4861 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     4921 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     4981 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     5041 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     5101 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     5161 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     5221 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     5281 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     5341 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     5401 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     5461 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     5521 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     5581 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     5641 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     5701 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     5761 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     5821 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     5881 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     5941 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     6001 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     6061 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     6121 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     6181 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     6241 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     6301 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     6361 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     6421 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     6481 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     6541 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     6601 cgaggggacc cttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct
     6661 ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt
     6721 ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc
     6781 ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa
     6841 cggaagccgc acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag
     6901 cgcgccgacc gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca
     6961 gcggccggga aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc
     7021 tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc
     7081 tcgttgaccg aatcaccgac ctctctcccc agggggtacc accggagctt accatgaccg
     7141 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     7201 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     7261 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     7321 aggtgtgggt cgcggacgac ggcgccgccg tggcggtctg gaccacgccg gagagcgtcg
     7381 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     7441 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     7501 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     7561 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     7621 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     7681 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagaattgt
     7741 agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     7801 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     7861 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     7921 cactttggcg ccggctcgag gggg
//
