LOCUS       sequence-232380-        8114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes codon optimized 3xFLAG-VpxMAC251 and puromycin resistance 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8114)
  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 8114)
  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-MAC251 vpx
FEATURES             Location/Qualifiers
     source          1..8114
                     /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             4220..4819
                     /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(4220..4239)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4716..4736
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4829..5417
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4882..4902)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5300..5311)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5935..6115
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6186..6774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6266..6285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6945..7736,7737..7805))
                     /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     7568..7587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7806..7910)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7985..8004)
                     /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 cgctatgagc gatccaaggg aacgaatccc gcccggaaat tctggtgaag
     3421 agaccatcgg tgaggcgttt gagtggctga accggactgt cgaggaaatc aacagggagg
     3481 ctgtgaacca tctgcccagg gaattgatat ttcaggtatg gcaacgatcc tgggagtatt
     3541 ggcatgacga acagggcatg tcacagagct acgtcaagta tcggtacttg tgtcttatgc
     3601 agaaagctct cttcatgcat tgcaaaaagg ggtgccgatg cttgggagaa ggtcatggtg
     3661 cgggggggtg gcgcccagga cccccacctc ctccaccccc tgggttggca taacccgggg
     3721 aattctcgac cttgactggc ggcgcgacct tgaggcctgc gttcgcctca gttgccccct
     3781 ctgtgcaatg gggagacgcg cctcatcgct tgacaacggc cgaagagccg ccgcgcttcc
     3841 gtctcccgcg tgcgcgcgcc atgctgccca cccccgttcc gcactgaccc tcccccgtgc
     3901 cccgcgtccc gtactgccgc cccgccccga gtcccatgcc gcagccaccg cgacggagcc
     3961 cgcaggcggg aacctgcctc cgcgcgttag cgcgcacgcg cgcctcatgt gtcgtcccca
     4021 tcagcgccgg cttccgtcta taggccagat gcactgtcac tctggcgaag tcgcagaccc
     4081 gattggccgg gacggaggcg cgagaccggg ttgcgggcgg ggccgaacgt ggtataaaag
     4141 gggcgggagg ccaggctcgt gccgttttgc agacgccacc gccgaggaaa accgtgtact
     4201 attagccacg cgtgccacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4261 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4321 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     4381 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     4441 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     4501 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     4561 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     4621 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     4681 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     4741 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     4801 ccgcaagccc ggtgcctgag cgatcgctaa tcaacctctg gattacaaaa tttgtgaaag
     4861 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4921 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4981 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5041 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5101 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5161 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5221 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5281 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5341 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5401 ttgggccgcc tccccgctta atcgcgtcga gacctagaaa aacatggagc aatcacaagt
     5461 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     5521 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     5581 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     5641 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag
     5701 aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta
     5761 gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta
     5821 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt
     5881 gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct
     5941 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6001 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6061 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc
     6121 cgtttcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7261 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7561 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7921 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     7981 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     8041 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     8101 acaattgcat gaag
//
