LOCUS       sequence-476616-        8851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral transfer plasmid encoding IL13 with GAP-Gly-Ser linker 
            and PDGFR TM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8851)
  AUTHORS   Dobson CS, Reich AN, Gaglione S, Smith BE, Kim EJ, Dong J, Ronsard 
            L, Okonkwo V, Lingwood D, Dougan M, Dougan SK, Birnbaum ME
  TITLE     Antigen identification and high-throughput interaction mapping by 
            reprogramming viral entry.
  JOURNAL   Nat Methods. 2022 Apr;19(4):449-460. doi: 
            10.1038/s41592-022-01436-z. Epub 2022 Apr 8.
  PUBMED    35396484
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: 
            "10.1038/s41592-022-01436-z"; journalName: "Nat Methods"; date: 
            "2022-04"; volume: "19"; issue: "4"; pages: "449-460"
COMMENT     Sequence Label: pHIV-IL13-GAPGlySer-PDGFR
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"
     promoter        2624..3802
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2855..3793
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3750..3770
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3829..3838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4231..4257
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(4418..4992,4993..4995,4996..5004)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4585..4602)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4812..4831
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5005..5007,5008..5010,5011..5724)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5005..5007,5008..5010,5011..5721)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5050..5071)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5311..5330)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5658..5679
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5725..6128
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     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 cgctctagcg tgaggctccg gtgcccgtca
     2641 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     2701 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     2761 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     2821 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
     2881 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
     2941 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
     3001 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
     3061 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
     3121 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
     3181 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     3241 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     3301 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     3361 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     3421 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     3481 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     3541 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     3601 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     3661 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     3721 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     3781 tcttccattt caggtgtcgt gagcggccgc tgagttaact attctagagc caccatggaa
     3841 actgacaccc tcttgctttg ggtgcttttg ttgtgggtac ccgggagtac gggtggtccg
     3901 gtccccccta gtacagcact tcgcgagctt atagaggaat tggtgaacat cacccagaac
     3961 caaaaggcgc ctctctgcaa cggttcaatg gtttggagta taaacttgac tgccggtatg
     4021 tattgcgcgg cgttggaatc actcatcaac gtatcaggtt gtagcgccat cgagaaaacg
     4081 caaagaatgc tttccggttt ctgtccccac aaggtaagcg cgggacaatt cagtagtttg
     4141 catgtacggg acactaaaat cgaggttgcc cagtttgtca aagaccttct gcttcatttg
     4201 aagaaactgt ttcgagaagg gcgattcaac tatccttacg atgtcccgga ctatgctggc
     4261 gcgccaggat ctggcggagg tggaagcgga ggcggtggat ctgctagcgt ggtggtgatc
     4321 tcagccatcc tggccctggt ggtgctcacc atcatctccc ttatcatcct catcatgctt
     4381 tggcagaaga agccacgttg agaattctga ggatccgccc ctctccctcc ccccccccta
     4441 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     4501 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     4561 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     4621 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     4681 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     4741 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     4801 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     4861 taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt
     4921 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     4981 cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg
     5041 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     5101 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     5161 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     5221 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     5281 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     5341 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     5401 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     5461 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     5521 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     5581 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     5641 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     5701 ggcatggacg agctgtacaa gtgaaatcaa cctctggatt acaaaatttg tgaaagattg
     5761 actgatattc ttaactatgt tgctcctttt acgctgtgtg gatatgctgc tttaatgcct
     5821 ctgtatcatg ctattgcttc ccgtacggct ttcgttttct cctccttgta taaatcctgg
     5881 ttgctgtctc tttatgagga gttgtggccc gttgtccgtc aacgtggcgt ggtgtgctct
     5941 gtgtttgctg acgcaacccc cactggctgg ggcattgcca ccacctgtca actcctttct
     6001 gggactttcg ctttccccct cccgatcgcc acggcagaac tcatcgccgc ctgccttgcc
     6061 cgctgctgga caggggctag gttgctgggc actgataatt ccgtggtgtt gtcggggaag
     6121 ctgacgtcct ttccatggct gctcgcctgt gttgccaact ggatcctgcg cgggacgtcc
     6181 ttctgctacg tcccttcggc tctcaatcca gcggacctcc cttcccgagg ccttctgccg
     6241 gttctgcggc ctctcccgcg tcttcgcttt cggcctccga cgagtcggat ctccctttgg
     6301 gccgcctccc cgcaagtaaa gcggccgcat 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
//
