LOCUS       sequence-232383-        8086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes codon optimized 3xFLAG-SIVMND2 Vpx (AY159322) and puromycin 
            resistance protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8086)
  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 8086)
  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-MND2 vpx
FEATURES             Location/Qualifiers
     source          1..8086
                     /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             4181..4780
                     /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(4181..4200)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4677..4697
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4790..5378
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4843..4863)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5261..5272)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5896..6076
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6158..6746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6238..6257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6917..7708,7709..7777))
                     /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     7540..7559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7778..7882)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7957..7976)
                     /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 cgctatggcg gaaagggctc ctgaagcccc agagggcgcg ggggaagttg
     3421 ggcttgagca gtggctggag acttctctcg aacgcatcaa tagagaggcg agactccact
     3481 tccatcctga gttccttttt aggctttgga atacgtgtgt cgagcactgg cacgataggc
     3541 accagcggag tttggattat gctaaatata gatatctcct gttgatgcac aaagcgatgt
     3601 acacccatat gcaacaaggg tgtccatgtc gcaatggcag acctcgtgga ccacccccac
     3661 ccggaatggc ctaacccggg gaattctcga ccttgactgg cggcgcgacc ttgaggcctg
     3721 cgttcgcctc agttgccccc tctgtgcaat ggggagacgc gcctcatcgc ttgacaacgg
     3781 ccgaagagcc gccgcgcttc cgtctcccgc gtgcgcgcgc catgctgccc acccccgttc
     3841 cgcactgacc ctcccccgtg ccccgcgtcc cgtactgccg ccccgccccg agtcccatgc
     3901 cgcagccacc gcgacggagc ccgcaggcgg gaacctgcct ccgcgcgtta gcgcgcacgc
     3961 gcgcctcatg tgtcgtcccc atcagcgccg gcttccgtct ataggccaga tgcactgtca
     4021 ctctggcgaa gtcgcagacc cgattggccg ggacggaggc gcgagaccgg gttgcgggcg
     4081 gggccgaacg tggtataaaa ggggcgggag gccaggctcg tgccgttttg cagacgccac
     4141 cgccgaggaa aaccgtgtac tattagccac gcgtgccacc atgaccgagt acaagcccac
     4201 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     4261 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     4321 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4381 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4441 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4501 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4561 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4621 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4681 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4741 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gcgatcgcta atcaacctct
     4801 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4861 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4921 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4981 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5041 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5101 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5161 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     5221 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5281 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5341 tcagacgagt cggatctccc tttgggccgc ctccccgctt aatcgcgtcg agacctagaa
     5401 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
     5461 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
     5521 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     5581 gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa
     5641 ggctacttcc ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc
     5701 tttggatggt gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa
     5761 ggagagaaca cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
     5821 gaagtattag agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg
     5881 catccggact gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     5941 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6001 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6061 tggaaaatct ctagcagggc ccgtttcatg tgagcaaaag gccagcaaaa ggccagcaaa
     6121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7921 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     7981 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     8041 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaag
//
