LOCUS       sequence-401801-        7744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HELLO neoantigen in tumor cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7744)
  AUTHORS   Cui C, Wang J, Fagerberg E, Chen PM, Connolly KA, Damo M, Cheung JF,
            Mao T, Askari AS, Chen S, Fitzgerald B, Foster GG, Eisenbarth SC, 
            Zhao H, Craft J, Joshi NS
  TITLE     Neoantigen-driven B cell and CD4 T follicular helper cell 
            collaboration promotes anti-tumor CD8 T cell responses.
  JOURNAL   Cell. 2021 Dec 9;184(25):6101-6118.e13. doi: 
            10.1016/j.cell.2021.11.007. Epub 2021 Nov 30.
  PUBMED    34852236
REFERENCE   2  (bases 1 to 7744)
  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.cell.2021.11.007"; journalName: "Cell"; date: "2021-12-9-
            9"; volume: "184"; issue: "25"; pages: "6101-6118.e13"
COMMENT     Sequence Label: pGK-HELLO LV
FEATURES             Location/Qualifiers
     source          1..7744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1683..1916
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2101..2145
                     /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    2443..2560
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     complement(2696..2719)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3093..3112
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3707..3730
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3791..3817
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3827..3880
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3881..4576
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(4014..4032)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4282..4301
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4632..5220
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4685..4705)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5103..5114)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5223..5239)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5224..5240)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5749..5929
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5991..6579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6071..6090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6750..7541,7542..7610))
                     /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     7373..7392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7611..7715)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gggtcgacta gtgatcagtg tgagggagtg
     2641 taaagctggt ttattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg
     2701 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca
     2761 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
     2821 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
     2881 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca
     2941 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg
     3001 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg
     3061 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc
     3121 tgccgcgctg ttctcctctt cctcatctcc gggactaact cgaacgctag ctgtgcgatc
     3181 gtttaccatg cgatctttgt tgatcctggt tttgtgtttt ttgcccctgg ctgctctcgg
     3241 gaaagtattc ggccggtgcg agctggctgc tgctatgaag cgacatgggt tggacaatta
     3301 tagaggctat agcctcggaa actgggtgtg cgccgcaaaa ttcgagagta atttcaatac
     3361 ccaggctaca aatagaaata cagatggttc aacagactat ggtattctcc agatcaatag
     3421 caggtggtgg tgtaacgacg gccgaacacc cggttctcgc aacctctgta acataccatg
     3481 ttcagccctt ctctctagcg atataactgc atccgttaat tgcgcaaaga aaatagtcag
     3541 tgatgggaat ggaatgaacg cttgggtcgc atggaggaac cgatgcaagg gtactgacgt
     3601 gcaggcttgg attcgaggct gtaggctggg tggaggcggt tcaggcggag gtggctctgg
     3661 cggtggcgga tcgaaagctg tgtacaactt tgcaacatgc gggatcgact acaaggatga
     3721 cgatgacaag ggtttgaacg gccctgacat ttacaaggga gtttatcagt ttaaatccgt
     3781 tgagtttgac tacccatacg atgttccaga ttacgctgga tctggagagg gcagaggaag
     3841 tctgctaaca tgcggtgacg tcgaggagaa tcctggccca atggtcagca agggcgaggc
     3901 agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga
     3961 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4021 gaaggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat
     4081 gtacggctcc agggccttca tcaagcaccc cgccgacatc cccgactact ataagcagtc
     4141 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac
     4201 cgtgacccag gacacctccc tggaggacgg caccctgatc tacaaggtca agctccgcgg
     4261 caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc
     4321 caccgagcgg ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg
     4381 cctgaaggac ggcggccgct acctggcgga cttcaagacc acctacaagg ccaagaagcc
     4441 cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg gacatcacct cccacaacga
     4501 ggactacacc gtggtggaac agtacgaacg ctccgagggc cgccactcca ccggcggcat
     4561 ggacgagctg tacaagtgaa aacgttgttg tttggggttg aattactctt ctagaccgcg
     4621 gcgcgcccga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4681 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4741 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4801 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4861 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4921 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4981 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     5041 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5101 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5161 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5221 atcgataccg tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat
     5281 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt
     5341 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     5401 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     5461 gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac
     5521 acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca
     5581 gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct
     5641 gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc
     5701 cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg
     5761 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5821 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5881 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag catgtgagca
     5941 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6001 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6061 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6121 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6181 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6241 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6301 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6361 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6421 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6481 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6541 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6601 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6661 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6721 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6781 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6841 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6901 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6961 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7021 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7081 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7141 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7201 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7261 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7321 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7381 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7441 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7501 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7561 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7621 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7681 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7741 gacg
//
