LOCUS       sequence-178816-        8247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of human codon optimized IL2 with 
            puromycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8247)
  AUTHORS   McCauley SM, Kim K, Nowosielska A, Dauphin A, Yurkovetskiy L, Diehl 
            WE, Luban J
  TITLE     Intron-containing RNA from the HIV-1 provirus activates type I 
            interferon and inflammatory cytokines.
  JOURNAL   Nat Commun. 2018 Dec 13;9(1):5305. doi: 10.1038/s41467-018-07753-2.
  PUBMED    30546110
REFERENCE   2  (bases 1 to 8247)
  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: "Nat 
            Commun."; date: "2018-12-13"; volume: "9(1):5305. doi"; pages: " 
            10.1038/s41467-018-07753-2"
COMMENT     Sequence Label: pAIP hIL2 co
FEATURES             Location/Qualifiers
     source          1..8247
                     /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"
     primer_bind     complement(240..260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        245..574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      425..560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     487..506
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        586..965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        967..1165
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1120..1140
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1183..1363
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1410..1535
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2032..2265
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2450..2494
                     /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    2792..2909
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3046..3453
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     sig_peptide     3480..3539
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     misc_feature    3950..4523
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4117..4134)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4344..4363
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4529..5128
                     /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(4529..4548)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5025..5045
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5138..5726
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5191..5211)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5609..5620)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6244..6424
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6495..7083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6575..6594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7254..8045,8046..8114))
                     /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     7877..7896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8115..8219)
                     /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 acgcgctgtg
      241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      481 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      541 aggaggcttt tttggaggcc taggcttttg caaaaagctc gcgttgacat tgattattga
      601 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      661 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      721 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      781 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      841 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      901 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      961 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1021 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1081 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1141 tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac
     1201 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     1261 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     1321 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     1381 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     1441 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     1501 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     1561 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     1621 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     1681 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     1741 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     1801 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     1861 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     1921 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     1981 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     2041 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     2101 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     2161 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     2221 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     2281 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     2341 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     2401 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     2461 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     2521 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     2581 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     2641 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     2701 agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat
     2761 tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     2821 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     2881 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag
     2941 tttggttaat taactgcagc cccgataaaa taaaagattt tatttagtct ccagaaaaag
     3001 gggggaatga aagaccccac ctgtaggttt ggcaagctag ctgcagtaac gccattttgc
     3061 aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc gggtacatga
     3121 aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc cccggcccgg
     3181 ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga acagatggtc
     3241 cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt ccaggctccc
     3301 ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct gcttctcgct
     3361 tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc ctcactcggc
     3421 gcgccagtcc tccgacagac tgagtcgccc gggggtctag aagcgctgga tccgccacca
     3481 tgtatagaat gcagctgctg tcctgtatcg cactgtcact ggctctggtc actaactccg
     3541 cacctacttc ctcctccaca aagaaaaccc agctgcagct ggaacacctg ctgctggacc
     3601 tgcagatgat tctgaacggc atcaacaatt acaagaatcc caaactgaca aggatgctga
     3661 ctttcaagtt ttatatgcct aagaaagcaa cagagctgaa gcacctgcag tgcctggagg
     3721 aagagctgaa accactggaa gaggtgctga acctggccca gagcaagaat ttccatctgc
     3781 ggcccagaga cctgatctcc aacattaatg tgatcgtcct ggaactgaaa gggtctgaga
     3841 ccaccttcat gtgcgaatac gccgatgaga ctgctaccat tgtggagttt ctgaaccgat
     3901 ggattacttt ttgccagagc attattagca ccctgacctg agaattccgc ccctctccct
     3961 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     4021 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggtc
     4081 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     4141 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     4201 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     4261 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     4321 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     4381 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     4441 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     4501 tcctttgaaa aacacgatga taaccggtat gaccgagtac aagcccacgg tgcgcctcgc
     4561 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4621 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4681 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4741 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4801 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4861 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4921 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4981 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5041 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5101 gtgcatgacc cgcaagcccg gtgcctgatt aatcgctaat caacctctgg attacaaaat
     5161 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5221 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5281 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5341 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5401 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5461 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5521 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     5581 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5641 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5701 gatctccctt tgggccgcct ccccgcttaa tcgcgtcgag acctagaaaa acatggagca
     5761 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     5821 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     5881 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5941 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     6001 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc
     6061 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc
     6121 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag
     6181 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt
     6241 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6301 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6361 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6421 agcagggccc gtttcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6901 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7381 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8221 acatttcccc gaaaagtgcc acctgac
//
