LOCUS       sequence-232539-        8183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes codon optimized 3xFLAG SIVGSN-CN71 Vpr (AF468658) and 
            puromycin resistance protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8183)
  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 8183)
  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-GSN-CN71 vpr
FEATURES             Location/Qualifiers
     source          1..8183
                     /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             4289..4888
                     /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(4289..4308)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4785..4805
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4898..5486
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4951..4971)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5369..5380)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6004..6184
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6255..6843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6335..6354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7014..7805,7806..7874))
                     /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     7637..7656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7875..7979)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8054..8073)
                     /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 cgctatggag cgaaccccac ctagtcaccc gttgccgtgg ataagtcgca
     3421 gagtaccgac taccatggca gtagctcaaa gtgccatgtg ggagataaac gaggaagcgg
     3481 aaaagcactt tagtcgagaa gagcttagag ggatctggca tgacgtgact gaacttccag
     3541 ccgacccaga ttggacagtt gaccaagcag ctattgcgtg tgccatagat tatgttaggc
     3601 gggtccaaac cctgttgttc cgccattttc gggacggatg ctttcaccgc tataaccgca
     3661 tcgtcaggag gtatcccgtg atacggccat tgcggggaac tgccccacct gattctaaca
     3721 gcgtgcctca tgctgatcct gaacagccga gaagaccgag cagatacaga atggatgaat
     3781 aacccgggga attctcgacc ttgactggcg gcgcgacctt gaggcctgcg ttcgcctcag
     3841 ttgccccctc tgtgcaatgg ggagacgcgc ctcatcgctt gacaacggcc gaagagccgc
     3901 cgcgcttccg tctcccgcgt gcgcgcgcca tgctgcccac ccccgttccg cactgaccct
     3961 cccccgtgcc ccgcgtcccg tactgccgcc ccgccccgag tcccatgccg cagccaccgc
     4021 gacggagccc gcaggcggga acctgcctcc gcgcgttagc gcgcacgcgc gcctcatgtg
     4081 tcgtccccat cagcgccggc ttccgtctat aggccagatg cactgtcact ctggcgaagt
     4141 cgcagacccg attggccggg acggaggcgc gagaccgggt tgcgggcggg gccgaacgtg
     4201 gtataaaagg ggcgggaggc caggctcgtg ccgttttgca gacgccaccg ccgaggaaaa
     4261 ccgtgtacta ttagccacgc gtgccaccat gaccgagtac aagcccacgg tgcgcctcgc
     4321 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4381 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4441 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4501 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4561 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4621 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4681 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4741 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     4801 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4861 gtgcatgacc cgcaagcccg gtgcctgagc gatcgctaat caacctctgg attacaaaat
     4921 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4981 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5041 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5101 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5161 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5221 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5281 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     5341 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5401 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5461 gatctccctt tgggccgcct ccccgcttaa tcgcgtcgag acctagaaaa acatggagca
     5521 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     5581 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     5641 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5701 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     5761 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc
     5821 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc
     5881 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag
     5941 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt
     6001 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6061 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6121 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6181 agcagggccc gtttcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6241 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6301 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6361 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6421 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6481 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6541 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6601 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6661 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6721 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6781 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6841 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6901 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6961 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7021 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7081 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7141 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7201 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7261 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7321 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7381 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7441 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7501 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7561 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7621 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7681 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7741 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7801 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7861 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7921 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7981 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8041 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8101 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8161 gcttgaccga caattgcatg aag
//
