LOCUS       sequence-479012-        8920 bp    DNA     circular SYN 18-JUL-2025
DEFINITION  Lentiviral transfer plasmid encoding IL13 on tetrameric coiled coil.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8920)
  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 8920)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 18, 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-TetCoil
FEATURES             Location/Qualifiers
     source          1..8920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(313..357)
                     /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    542..775
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1272..1397
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1444..1624
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1642..1840)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1667..1687)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1842..2221
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2393..2412
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2487..2591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2592..2660,2661..3452)
                     /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(2810..2829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3623..4211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4112..4131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             4273..4453
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     4962..4978
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4963..4979
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5182..5585
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(join(5586..6299,6300..6302,6303..6305))
                     /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             complement(join(5589..6299,6300..6302,6303..6305))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5631..5652)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5980..5999
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6239..6260
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    join(6306..6314,6315..6317,6318..6892)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6479..6498)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6708..6725
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(7122..7148)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      7541..7550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7577..8755)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7586..8524
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(7609..7629)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(8818..8920,1..15)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ccttttcttt taaaattgtg gatgaatact gccatttgtc tgcagaattg gcgcacgcag
       61 tgccgatccg ttcactaatc gaatggatct gtctctgtct ctctctccac cttcttcttc
      121 tattccttcg ggcctgtcgg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
      181 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
      241 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
      301 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
      361 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
      421 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
      481 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
      541 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
      601 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
      661 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
      721 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
      781 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
      841 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
      901 tcctcctcca ggtctgaaga tcagcggccg gccgcttgct gtgcggtggt cttacttttg
      961 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     1021 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     1081 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     1141 ccaggattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     1201 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     1261 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     1321 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     1381 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     1441 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     1501 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     1561 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     1621 acccagtaca ggcaaaacgc gctgcttata tagacctccc accgtacacg cctaccgccc
     1681 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     1741 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     1801 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     1861 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     1921 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     1981 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     2041 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     2101 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     2161 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     2221 caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga
     2281 ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac
     2341 tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca
     2401 gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt
     2461 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2521 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2581 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2641 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2701 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2761 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2821 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2881 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2941 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3001 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3061 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3121 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3181 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3241 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3301 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3361 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3421 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3481 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3541 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4261 ttgctcacat gctgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt
     4321 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
     4381 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
     4441 accagagaga cccagtacag tccggatgca gctctcgggc catgtgatga aatgctaggc
     4501 ggctgtcaaa cctccactct aatacttctc tctccgggtc atccatccca tgcaggctca
     4561 cagggtgtaa caagcgggtg ttctctcctt cattggcttc ttctaccttc tcttgctcaa
     4621 ctggtactag cttgtagcac catccaaagg tcagtggata tctgatccct ggccctggtg
     4681 tgtagttctg ccaatcaggg aagtagcctt gtgtgtggta gatccacaga tcaaggatat
     4741 cttgtcttcg ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc
     4801 taagatctac agctgccttg taagtcattg gtcttaaagg tacctgaggt gtgactggaa
     4861 aacccacctc ctcctcctct tgtgcttcta gccaggcaca atcagcattg gtagctgctg
     4921 tattgctact tgtgattgct ccatgttttt ctaggtctcg atcgaggtcg acggtatcga
     4981 tgcggccgct ttacttgcgg ggaggcggcc caaagggaga tccgactcgt cggaggccga
     5041 aagcgaagac gcgggagagg ccgcagaacc ggcagaaggc ctcgggaagg gaggtccgct
     5101 ggattgagag ccgaagggac gtagcagaag gacgtcccgc gcaggatcca gttggcaaca
     5161 caggcgagca gccatggaaa ggacgtcagc ttccccgaca acaccacgga attatcagtg
     5221 cccagcaacc tagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttctgccgtg
     5281 gcgatcggga gggggaaagc gaaagtccca gaaaggagtt gacaggtggt ggcaatgccc
     5341 cagccagtgg gggttgcgtc agcaaacaca gagcacacca cgccacgttg acggacaacg
     5401 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaacgaaa
     5461 gccgtacggg aagcaatagc atgatacaga ggcattaaag cagcatatcc acacagcgta
     5521 aaaggagcaa catagttaag aatatcagtc aatctttcac aaattttgta atccagaggt
     5581 tgatttcact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     5641 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc
     5701 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     5761 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     5821 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     5881 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     5941 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     6001 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     6061 tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc
     6121 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     6181 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     6241 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     6301 accatggttg tggccatatt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt
     6361 cggggggcct agacgttttt ttaacctcga ctaaacacat gtaaagcatg tgtaccgagg
     6421 ccccagatca gatcccatac aatggggtac cttctgggca tccttcagcc ccttgttgaa
     6481 tacgcttgag gagagccatt tgactctttc cacaactatc caactcacaa cgtggcactg
     6541 gggttgtgcc gcctttgcag gtgtatctta tacacgtggc ttttggccgc agaggcacct
     6601 gtcgccaggt ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca
     6661 agaagcttcc agaggaactg cttccttcac gacattcaac agaccttgca ttcctttggc
     6721 gagaggggaa agacccctag gaatgctcgt caagaagaca gggccaggtt tccgggccct
     6781 cacattgcca aaagacggca atatggtgga aaataacata tagacaaacg cacaccggcc
     6841 ttattccaag cggcttcggc cagtaacgtt aggggggggg gagggagagg ggcggatcct
     6901 cagaattctc aacgtggctt cttctgccaa agcatgatga ggatgataag ggagatgatg
     6961 gtgagcacca ccagggccag gatggctgag atcaccacca ccttgccagc cccttgcttg
     7021 atagccgcca gctcccactt aattgccgcc agctcctttt tgattgctgc aagctcttgt
     7081 tttatcgccg ccaactcgcc gcttccacct ccgccgctag cagcatagtc cgggacatcg
     7141 taaggatagt tgaatcgccc ttctcgaaac agtttcttca aatgaagcag aaggtctttg
     7201 acaaactggg caacctcgat tttagtgtcc cgtacatgca aactactgaa ttgtcccgcg
     7261 cttaccttgt ggggacagaa accggaaagc attctttgcg ttttctcgat ggcactacaa
     7321 cctgatacgt tgatgagtga ttccaacgcc gcgcaataca taccggcagt caagtttata
     7381 ctccaaacca ttgaaccgtt gcagagaggc gccttttggt tctgggtgat gttcaccaat
     7441 tcctctataa gctcgcgaag tgctgtacta ggggggaccg gaccacccgt actcccgggt
     7501 acccacaaca aaagcaccca aagcaagagg gtgtcagttt ccatggtggc tctagaatag
     7561 ttaactcagc ggccgctcac gacacctgaa atggaagaaa aaaactttga accactgtct
     7621 gaggcttgag aatgaaccaa gatccaaact caaaaagggc aaattccaag gagaattaca
     7681 tcaagtgcca agctggccta acttcagtct ccacccactc agtgtgggga aactccatcg
     7741 cataaaaccc ctccccccaa cctaaagacg acgtactcca aaagctcgag aactaatcga
     7801 ggtgcctgga cggcgcccgg tactccgtgg agtcacatga agcgacggct gaggacggaa
     7861 aggccctttt cctttgtgtg ggtgactcac ccgcccgctc tcccgagcgc cgcgtcctcc
     7921 attttgagct ccctgcagca gggccgggaa gcggccatct ttccgctcac gcaactggtg
     7981 ccgaccgggc cagccttgcc gcccagggcg gggcgataca cggcggcgcg aggccaggca
     8041 ccagagcagg ccggccagct tgagactacc cccgtccgat tctcggtggc cgcgctcgca
     8101 ggccccgcct cgccgaacat gtgcgctggg acgcacgggc cccgtcgccg cccgcggccc
     8161 caaaaaccga aataccagtg tgcagatctt ggcccgcatt tacaagacta tcttgccaga
     8221 aaaaaagcgt cgcagcaggt catcaaaaat tttaaatggc tagagactta tcgaaagcag
     8281 cgagacaggc gcgaaggtgc caccagattc gcacgcggcg gccccagcgc ccaggccagg
     8341 cctcaactca agcacgaggc gaaggggctc cttaagcgca aggcctcgaa ctctcccacc
     8401 cacttccaac ccgaagctcg ggatcaagaa tcacgtactg cagccaggtg gaagtaattc
     8461 aaggcacgca agggccataa cccgtaaaga ggccaggccc gcgggaacca cacacggcac
     8521 ttacctgtgt tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac
     8581 ttatatacgg ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt
     8641 cccagtttac cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca
     8701 acttctcggg gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagcctcacg
     8761 ctagagcggg cccggtacta accaaactgg atctctgctg tccctgtaat aaacccgaaa
     8821 attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat
     8881 gtctactatt ctttcccctg cactgtaccc cccaatcccc
//
