LOCUS       sequence-232371-        9662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SFFV promoter expresses gag-gfp fusion with CypA promoter driving 
            expression of SIVMAC251 vpx.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9662)
  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 9662)
  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 gag-gfp/vpx
FEATURES             Location/Qualifiers
     source          1..9662
                     /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"
     regulatory      3289..3298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4809..4811,4812..4814,4815..5525)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    6377..6965
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6430..6450)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6848..6859)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7483..7663
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7734..8322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7814..7833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8493..9284,9285..9353))
                     /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     9116..9135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9354..9458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9533..9552)
                     /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 atggcggcaa
     3301 gggcaagcgt gctgagcggg ggggagctgg acaaatggga gaaaatccgg ctgagacctg
     3361 gaggcaagaa acagtataaa ctgaagcaca tcgtgtgggc tagcagagaa ctggagaggt
     3421 tcgcagtcaa ccccggcctg ctggaaacat ccgagggatg ccgacagatt ctggggcagc
     3481 tgcagccttc cctgcagact ggatccgagg aactgcgcag tctgtacaac acagtggcca
     3541 ccctgtattg cgtgcatcag cggattgacg tgaaagatac caaggaggct ctggacaaaa
     3601 tcgaggaaga gcagaataag tccaagaaaa aggcacagca ggcggccgca gatacaggga
     3661 acaatagcca ggtgtcccag aactacccaa tcgtccagaa tctgcaggga cagatggtgc
     3721 accaggccat ttctcccaga accctgaacg cttgggtgaa agtggtcgaa gagaaggcct
     3781 tcagtcctga agtcatccca atgttttctg ctctgagtga gggcgcaaca cctcaagatc
     3841 taaacacaat gctgaatact gtgggcgggc atcaggccgc tatgcagatg ctgaaggaaa
     3901 ctatcaatga agaggcagca gagtgggaca ggctgcaccc agtgcatgca ggaccaattg
     3961 cacctgggca gatgcgagaa cctagaggat cagatatcgc cggcaccaca agcacactgc
     4021 aggagcagat tgggtggatg actcacaacc cccctatacc ggtaggagaa atctacaaaa
     4081 ggtggatcat tctgggcctg aataagatcg tccgcatgta ttcacccact agcatcctgg
     4141 acattcgaca aggcccaaaa gagcccttcc gcgactacgt ggacagattc tacaagaccc
     4201 tgcgcgccga acaagcttca caggaggtga aaaactggat gactgagacc ctgctggtcc
     4261 agaacgctaa tcctgattgc aaaacaattc tgaaggccct ggggccagga gctactctgg
     4321 aagagatgat gaccgcatgt cagggagtgg gaggacctgg acacaaggcc agagtgctgg
     4381 ctgaggcaat gagccaggtc accaatccag ccacaatcat gattcagaag ggcaacttcc
     4441 ggaaccagcg caaaaccgtg aagtgcttta actgtggcaa agaggggcat atcgcaaaga
     4501 attgcagggc ccctcggaaa aagggatgct ggaaatgtgg aaaggaaggc caccagatga
     4561 aggactgtac tgagagacag gctaacttcc tgggcaaaat ttggccctcc cataagggga
     4621 gacccgggaa ttttctgcag agccggcctg agccaaccgc accacccgaa gagagtttca
     4681 ggtttggcga agagactacc acaccatcac agaaacagga acccatcgac aaggagctgt
     4741 atcctctggc aagtctgcga agcctgtttg gaagcgaccc ctcatctcag ggcggaagcg
     4801 gcggatccat ggtctcaaag ggggaagagc tgtttactgg ggtcgtgcct attctggtcg
     4861 aactggacgg ggatgtcaac ggccacaagt ttagtgtgag cggagaggga gaaggcgacg
     4921 ccacttacgg aaagctgacc ctgaaattca tttgcaccac aggcaagctg ccagtgccct
     4981 ggcctaccct ggtcactacc ctgacatacg gcgtccagtg ttttagtagg tatcctgacc
     5041 acatgaagca gcatgatttc tttaaatcag ctatgccaga gggatacgtg caggaacgca
     5101 ccattttctt taaggacgat ggcaactaca agactcgagc agaggtgaag ttcgaagggg
     5161 acaccctggt caaccggatc gagctgaagg gaatcgactt taaagaagat ggaaacatcc
     5221 tgggccacaa gctggagtac aactataatt ctcataacgt gtacatcatg gccgataagc
     5281 agaaaaacgg gattaaggtc aatttcaaaa tcagacacaa catcgaagac ggcagcgtgc
     5341 agctggctga tcattaccag cagaacacac caattggcga cgggccagtg ctgctgccag
     5401 ataatcacta tctgagcact cagtccgccc tgtctaagga ccctaacgag aaacgcgatc
     5461 atatggtgct gctggagttc gtcacagccg ctgggatcac tctgggaatg gacgagctgt
     5521 ataaatgaga attctcgacc ttgactggcg gcgcgacctt gaggcctgcg ttcgcctcag
     5581 ttgccccctc tgtgcaatgg ggagacgcgc ctcatcgctt gacaacggcc gaagagccgc
     5641 cgcgcttccg tctcccgcgt gcgcgcgcca tgctgcccac ccccgttccg cactgaccct
     5701 cccccgtgcc ccgcgtcccg tactgccgcc ccgccccgag tcccatgccg cagccaccgc
     5761 gacggagccc gcaggcggga acctgcctcc gcgcgttagc gcgcacgcgc gcctcatgtg
     5821 tcgtccccat cagcgccggc ttccgtctat aggccagatg cactgtcact ctggcgaagt
     5881 cgcagacccg attggccggg acggaggcgc gagaccgggt tgcgggcggg gccgaacgtg
     5941 gtataaaagg ggcgggaggc caggctcgtg ccgttttgca gacgccaccg ccgaggaaaa
     6001 ccgtgtacta ttagccacgc gtgccaccat gagcgaccca agagagagaa tcccaccagg
     6061 caatagcggc gaagaaacaa tcggagaggc ttttgaatgg ctgaacagga ccgtcgagga
     6121 aatcaaccgg gaggcagtga atcacctgcc tagagaactg attttccagg tctggcagcg
     6181 atcttgggag tactggcatg acgaacaggg gatgagccag tcctatgtga agtacagata
     6241 tctgtgcctg atgcagaaag ccctgtttat gcactgtaag aaaggctgca ggtgtctggg
     6301 agagggccat ggggccggcg ggtggcgccc cggaccccct ccaccccctc cacccggcct
     6361 ggcttgagcg atcgctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     6421 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     6481 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6541 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6601 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6661 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6721 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     6781 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6841 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6901 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6961 cccgcttaat cgcgtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     7021 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     7081 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     7141 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
     7201 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
     7261 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     7321 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
     7381 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
     7441 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
     7501 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     7561 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     7621 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttcatgtga
     7681 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7741 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7801 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7861 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7921 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7981 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     8041 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     8101 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     8161 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8221 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     8281 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     8341 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     8401 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8461 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8521 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8581 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8641 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8701 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8761 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8821 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8881 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8941 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     9001 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     9061 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     9121 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     9181 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     9241 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     9301 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     9361 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     9421 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9481 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     9541 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     9601 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     9661 ag
//
