LOCUS       sequence-232480-        8104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes codon optimized 3xFLAG-SIVAGI Vpx (HM803690) and puromycin 
            resistance protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8104)
  AUTHORS   Yurkovetskiy L, Guney MH, Kim K, Goh SL, McCauley S, Dauphin A, 
            Diehl WE, Luban J
  TITLE     Primate immunodeficiency virus proteins Vpx and Vpr counteract 
            transcriptional repression of proviruses by the HUSH complex.
  JOURNAL   Nat Microbiol. 2018 Dec;3(12):1354-1361. doi: 
            10.1038/s41564-018-0256-x. Epub 2018 Oct 8.
  PUBMED    30297740
REFERENCE   2  (bases 1 to 8104)
  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/s41564-018-0256-x"; journalName: "Nat Microbiol"; date: 
            "2018-12"; volume: "3"; issue: "12"; pages: "1354-1361"
COMMENT     Sequence Label: pscALPS-SIV-AGI vpx
FEATURES             Location/Qualifiers
     source          1..8104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..84)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        69..398
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      249..384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     311..330
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        410..789
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        791..989
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     944..964
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1007..1187
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1234..1359
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1856..2089
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2273..2317
                     /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    2615..2732
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2869..3276
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3285..3294
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3294..3365
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             4199..4798
                     /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(4199..4218)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4695..4715
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4808..5396
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4861..4881)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5279..5290)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5914..6094
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6176..6764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6256..6275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6935..7726,7727..7795))
                     /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     7558..7577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7796..7900)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7975..7994)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc
       61 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
      121 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
      181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
      241 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
      301 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
      361 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgcgttg acattgatta
      421 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      481 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      541 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      601 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      661 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      721 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      781 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      841 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      901 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      961 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt
     1021 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     1081 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     1141 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     1201 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     1261 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     1321 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     1381 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     1441 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     1501 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     1561 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     1621 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     1681 accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg
     1741 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1801 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1861 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1921 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1981 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     2041 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     2101 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     2161 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaattaacaa
     2221 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     2281 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     2341 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     2401 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     2461 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     2521 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc
     2581 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     2641 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2701 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     2761 cagtttggtt aattaactgc agccccgata aaataaaaga ttttatttag tctccagaaa
     2821 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagctgcagt aacgccattt
     2881 tgcaaggcat ggaaaaatac caaaccaaga atagagaagt tcagatcaag ggcgggtaca
     2941 tgaaaatagc taacgttggg ccaaacagga tatctgcggt gagcagtttc ggccccggcc
     3001 cggggccaag aacagatggt caccgcagtt tcggccccgg cccgaggcca agaacagatg
     3061 gtccccagat atggcccaac cctcagcagt ttcttaagac ccatcagatg tttccaggct
     3121 cccccaagga cctgaaatga ccctgcgcct tatttgaatt aaccaatcag cctgcttctc
     3181 gcttctgttc gcgcgcttct gcttcccgag ctctataaaa gagctcacaa cccctcactc
     3241 ggcgcgccag tcctccgaca gactgagtcg cccgggggtc tagagccgcc atggattata
     3301 aagatgacga tgacaaagac tataaagacg atgacgacaa agattacaag gacgacgacg
     3361 acaaagcggc cgctatggct gaacgggcac cagaaggacc cacgggtgcg ggcgaagtag
     3421 aatttcaacc gtggctggca cgaatgctgt atgaaatcaa ccaagaggcc cggctccatt
     3481 tcccaggaga gttcattttt cacctgtggc gaacttgcgt tgagtactgg cacgatgaac
     3541 aagggagaac actggaatat gccggatacc gctacctttt gttgatgcag aaagcactct
     3601 ttactcatat gaggagtggg tgcaagttgc gacatggcga ccccccaagg cagcgaggag
     3661 aacgggtacc aatattgccg ggtatgcagt aacccgggga attctcgacc ttgactggcg
     3721 gcgcgacctt gaggcctgcg ttcgcctcag ttgccccctc tgtgcaatgg ggagacgcgc
     3781 ctcatcgctt gacaacggcc gaagagccgc cgcgcttccg tctcccgcgt gcgcgcgcca
     3841 tgctgcccac ccccgttccg cactgaccct cccccgtgcc ccgcgtcccg tactgccgcc
     3901 ccgccccgag tcccatgccg cagccaccgc gacggagccc gcaggcggga acctgcctcc
     3961 gcgcgttagc gcgcacgcgc gcctcatgtg tcgtccccat cagcgccggc ttccgtctat
     4021 aggccagatg cactgtcact ctggcgaagt cgcagacccg attggccggg acggaggcgc
     4081 gagaccgggt tgcgggcggg gccgaacgtg gtataaaagg ggcgggaggc caggctcgtg
     4141 ccgttttgca gacgccaccg ccgaggaaaa ccgtgtacta ttagccacgc gtgccaccat
     4201 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     4261 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     4321 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     4381 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     4441 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     4501 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     4561 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     4621 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     4681 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     4741 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagc
     4801 gatcgctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4861 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4921 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4981 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5041 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5101 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5161 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     5221 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5281 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5341 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcttaa
     5401 tcgcgtcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     5461 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     5521 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     5581 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     5641 tggatctacc acacacaagg ctacttccct gattggcaga actacacacc agggccaggg
     5701 atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag
     5761 gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg
     5821 atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat
     5881 cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga
     5941 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6001 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6061 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttcatgtg agcaaaaggc
     6121 cagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6181 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6241 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6301 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6361 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6421 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6481 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6541 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6601 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6661 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6721 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6781 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6841 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6901 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6961 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7021 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7081 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7141 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7201 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7261 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7321 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7381 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7441 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7501 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7561 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7621 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7681 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7741 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7801 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7861 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7921 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     7981 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     8041 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     8101 gaag
//
