LOCUS       sequence-401802-        8851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generates lentiviral vector for autochthonous KP-HELLO tumor 
            initiation by intratracheal administration to KP mice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8851)
  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 8851)
  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-T2A-Cre LV
FEATURES             Location/Qualifiers
     source          1..8851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     complement(2697..2720)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3094..3113
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3708..3731
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3792..3818
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3828..3881
                     /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             3882..4577
                     /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(4015..4033)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4283..4302
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4578..4631
                     /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             4638..4658
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5740..6328
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5793..5813)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6211..6222)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6331..6347)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6332..6348)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6857..7037
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7099..7687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7179..7198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7858..8649,8650..8718))
                     /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     8481..8500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8719..8823)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cgggtcgact agtgatcagt gtgagggagt
     2641 gtaaagctgg tttattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     2701 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     2761 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     2821 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     2881 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     2941 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     3001 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     3061 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     3121 ctgccgcgct gttctcctct tcctcatctc cgggactaac tcgaacgcta gctgtgcgat
     3181 cgtttaccat gcgatctttg ttgatcctgg ttttgtgttt tttgcccctg gctgctctcg
     3241 ggaaagtatt cggccggtgc gagctggctg ctgctatgaa gcgacatggg ttggacaatt
     3301 atagaggcta tagcctcgga aactgggtgt gcgccgcaaa attcgagagt aatttcaata
     3361 cccaggctac aaatagaaat acagatggtt caacagacta tggtattctc cagatcaata
     3421 gcaggtggtg gtgtaacgac ggccgaacac ccggttctcg caacctctgt aacataccat
     3481 gttcagccct tctctctagc gatataactg catccgttaa ttgcgcaaag aaaatagtca
     3541 gtgatgggaa tggaatgaac gcttgggtcg catggaggaa ccgatgcaag ggtactgacg
     3601 tgcaggcttg gattcgaggc tgtaggctgg gtggaggcgg ttcaggcgga ggtggctctg
     3661 gcggtggcgg atcgaaagct gtgtacaact ttgcaacatg cgggatcgac tacaaggatg
     3721 acgatgacaa gggtttgaac ggccctgaca tttacaaggg agtttatcag tttaaatccg
     3781 ttgagtttga ctacccatac gatgttccag attacgctgg atctggagag ggcagaggaa
     3841 gtctgctaac atgcggtgac gtcgaggaga atcctggccc aatggtcagc aagggcgagg
     3901 cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg
     3961 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     4021 tgaaggtgac caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca
     4081 tgtacggctc cagggccttc atcaagcacc ccgccgacat ccccgactac tataagcagt
     4141 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga
     4201 ccgtgaccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtc aagctccgcg
     4261 gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt
     4321 ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc
     4381 gcctgaagga cggcggccgc tacctggcgg acttcaagac cacctacaag gccaagaagc
     4441 ccgtgcagat gcccggcgcc tacaacgtcg accgcaagtt ggacatcacc tcccacaacg
     4501 aggactacac cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca
     4561 tggacgagct gtacaaggag ggacgaggtt ctttgctgac ttgcggcgat gtggaagaaa
     4621 acccggggcc aggcatgccc aagaagaagc gcaaagtgtc taacctcctg acagtgcacc
     4681 aaaacctccc ggcactgcca gtggatgcta cttctgacga agtgcgcaaa aatcttatgg
     4741 acatgttccg cgaccggcaa gcttttagcg aacatacttg gaagatgctg ttgagtgtct
     4801 gccgctcttg ggccgcttgg tgcaaactca acaaccggaa gtggtttccc gctgagcctg
     4861 aggatgttcg ggactatctc ctgtaccttc aagcaagagg cctcgcagtg aagactatcc
     4921 aacagcacct gggccagctg aacatgctgc atcggagaag cggattgccc cgaccctccg
     4981 atagcaatgc agttagtttg gtaatgagga gaatcaggaa agaaaatgtg gatgctggag
     5041 agagggccaa acaggctttg gcattcgaaa gaaccgattt cgaccaggtc cgtagcctga
     5101 tggagaactc agataggtgc caagatataa gaaacctggc ctttcttggc atcgcttata
     5161 acacactgct gcgcattgcc gagatcgccc gcatccgggt gaaggacatc agcagaactg
     5221 atggcggcag gatgctcatc catatcggcc ggactaaaac actggtctcc accgccgggg
     5281 tcgagaaggc tctgagcctc ggcgttacca agctcgtgga aagatggatc agcgtgtctg
     5341 gtgtggccga cgacccaaac aactacctgt tttgccgagt gagaaaaaat ggcgtggcag
     5401 ccccttctgc aacttctcaa ctgagtactc gcgcactcga agggatcttt gaggctactc
     5461 accggctgat ctacggtgct aaggacgata gcggacagcg gtatctggcc tggtccggac
     5521 acagcgcacg agtcggcgct gcccgggaca tggctagggc tggggtgtcc attcccgaga
     5581 ttatgcaagc tgggggctgg actaacgtta atatcgtgat gaactacatc agaaatcttg
     5641 actccgaaac aggcgctatg gtgcggctgt tggaagacgg agactaaaaa cgttgttgtt
     5701 tggggttgaa ttactcttct agaccgcggc gcgcccgata atcaacctct ggattacaaa
     5761 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5821 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5881 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5941 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     6001 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     6061 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     6121 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     6181 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     6241 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     6301 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgatc gagacctaga
     6361 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
     6421 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc
     6481 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     6541 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca
     6601 aggctacttc cctgattggc agaactacac accagggcca gggatcagat atccactgac
     6661 ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga
     6721 aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
     6781 agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct
     6841 gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6901 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6961 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     7021 gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7261 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7501 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8701 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8761 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8821 gcgcacattt ccccgaaaag tgccacctga c
//
