LOCUS       sequence-328006-        7945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CMV US20 (29-37) fused to IL-2 signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7945)
  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 7945)
  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: VYAIFIFQL_epitope
FEATURES             Location/Qualifiers
     source          1..7945
                     /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    756..1344
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(809..829)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1227..1238)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1416..1649
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1727..1848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1764..1783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1818..1837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1888..2023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2043..2062)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2044..2062)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2072..2089)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2072..2088)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2081..2103)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2230..2685
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2317..2336)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2527..2548
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2711..2815
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2816..2884,2885..3676)
                     /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(3034..3053)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3847..4435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4336..4355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4589..4606
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4723..4744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4759..4764,4765..4782,4783..4789)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4797..4813
                     /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     4802..4824
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4821..4837
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4821..4837
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4856..4876
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4858..4876
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4904..5130
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5131..5311
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5358..5483
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5976..6209
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6394..6438
                     /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        6613..7113
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7032..7050
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7135..7734
                     /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(7135..7154)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7631..7651
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7776..7893
                     /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 ttgtcgccac catgtacagg atgcaactcc tgtcttgcat tgcactaagt cttgcacttg
      661 tcacgaattc agtgtacgcc atcttcatct tccagctgtg aattcaaagt ggtttagtaa
      721 tgagctagcg ctaaccggtg gcgcgttaag tcgacaatca acctctggat tacaaaattt
      781 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
      841 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
      901 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
      961 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     1021 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     1081 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     1141 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     1201 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     1261 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     1321 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
     1381 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     1441 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     1501 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     1561 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     1621 cttttagtca gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat
     1681 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     1741 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1801 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     1861 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     1921 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     1981 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     2041 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     2101 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2161 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2221 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2281 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2341 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2401 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2461 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2521 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2581 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2641 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     2701 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2761 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2821 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2881 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2941 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3001 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3061 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3121 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3181 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3241 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3301 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3361 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3421 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3481 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3541 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3601 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3661 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3721 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3781 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3841 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3901 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3961 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4021 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4081 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4141 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4201 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4261 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4321 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4381 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4441 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4501 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4561 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4621 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4681 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4741 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4801 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     4861 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     4921 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     4981 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     5041 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     5101 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     5161 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5221 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     5281 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     5341 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     5401 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     5461 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     5521 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     5581 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     5641 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     5701 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     5761 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     5821 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     5881 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     5941 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     6001 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     6061 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     6121 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     6181 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     6241 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     6301 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     6361 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     6421 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     6481 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     6541 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     6601 ccgaggggac ccttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc
     6661 tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg
     6721 tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt
     6781 cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa
     6841 acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca
     6901 gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc
     6961 agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc
     7021 ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc
     7081 ctcgttgacc gaatcaccga cctctctccc cagggggtac caccggagct taccatgacc
     7141 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     7201 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     7261 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     7321 aaggtgtggg tcgcggacga cggcgccgcc gtggcggtct ggaccacgcc ggagagcgtc
     7381 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     7441 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     7501 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     7561 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     7621 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     7681 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgagaattc
     7741 tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     7801 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     7861 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     7921 ccactttggc gccggctcga ggggg
//
