LOCUS       sequence-145997-        8216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of human codon optimized GMCSF with
            puromycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8216)
  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 8216)
  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 hGMCSF co
FEATURES             Location/Qualifiers
     source          1..8216
                     /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        582..961
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        963..1161
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1116..1136
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1179..1359
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1406..1531
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2028..2261
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2446..2490
                     /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    2788..2905
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3042..3449
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    3919..4492
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4086..4103)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4313..4332
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4498..5097
                     /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(4498..4517)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4994..5014
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5107..5695
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5160..5180)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5578..5589)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6213..6393
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6464..7052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6544..6563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7223..8014,8015..8083))
                     /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     7846..7865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8084..8188)
                     /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 caaaaagctt tgacattgat tattgactag
      601 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      661 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      721 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      781 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      841 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      901 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      961 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1021 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1081 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1141 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
     1201 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     1261 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     1321 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     1381 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     1441 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     1501 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     1561 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     1621 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     1681 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1741 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1801 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1861 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     1921 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     1981 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     2041 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     2101 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     2161 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     2221 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     2281 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     2341 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     2401 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     2461 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     2521 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     2581 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     2641 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     2701 agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg
     2761 cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     2821 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     2881 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
     2941 gttaattaac tgcagccccg ataaaataaa agattttatt tagtctccag aaaaaggggg
     3001 gaatgaaaga ccccacctgt aggtttggca agctagctgc agtaacgcca ttttgcaagg
     3061 catggaaaaa taccaaacca agaatagaga agttcagatc aagggcgggt acatgaaaat
     3121 agctaacgtt gggccaaaca ggatatctgc ggtgagcagt ttcggccccg gcccggggcc
     3181 aagaacagat ggtcaccgca gtttcggccc cggcccgagg ccaagaacag atggtcccca
     3241 gatatggccc aaccctcagc agtttcttaa gacccatcag atgtttccag gctcccccaa
     3301 ggacctgaaa tgaccctgcg ccttatttga attaaccaat cagcctgctt ctcgcttctg
     3361 ttcgcgcgct tctgcttccc gagctctata aaagagctca caacccctca ctcggcgcgc
     3421 cagtcctccg acagactgag tcgcccgggg gtctagaagc gctggatccg ccaccatgtg
     3481 gctgcagtcc ctgctgctgc tgggcaccgt cgcctgttct atctccgctc ccgcacgctc
     3541 tccaagtccc tcaactcagc cttgggaaca cgtgaacgca atccaggagg cccggagact
     3601 gctgaatctg agccgagaca ccgccgctga gatgaacgaa acagtggagg tcatttctga
     3661 aatgttcgat ctgcaggagc ccacttgcct gcagaccagg ctggaactgt acaagcaggg
     3721 cctgcgcggg tctctgacta agctgaaagg ccctctgaca atgatggcta gtcactataa
     3781 acagcattgc ccccctaccc cagagacaag ctgtgccaca cagatcatta ctttcgaatc
     3841 ctttaaggag aatctgaaag acttcctgct ggtcatccca tttgattgtt gggaacccgt
     3901 ccaggagtga gaattccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
     3961 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     4021 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     4081 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     4141 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     4201 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     4261 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     4321 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     4381 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     4441 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aaccggtatg
     4501 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     4561 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     4621 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     4681 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     4741 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     4801 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     4861 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     4921 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     4981 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5041 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatta
     5101 atcgctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5161 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5221 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5281 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5341 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5401 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5461 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     5521 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     5581 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5641 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcttaat
     5701 cgcgtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
     5761 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
     5821 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5881 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
     5941 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
     6001 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
     6061 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
     6121 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
     6181 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
     6241 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6301 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6361 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttcatgtga gcaaaaggcc
     6421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6901 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
