LOCUS       sequence-251145-        7743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes puromycin resistance protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7743)
  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 7743)
  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_puro
FEATURES             Location/Qualifiers
     source          1..7743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..441
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        443..641
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     596..616
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             659..839
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    886..1011
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1508..1741
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1925..1969
                     /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    2267..2384
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2521..2928
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3501..4100
                     /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(3501..3520)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3997..4017
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4110..4698
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4163..4183)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4581..4592)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5216..5396
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5467..6055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5547..5566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6226..7017,7018..7086))
                     /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     6849..6868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7087..7191)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7266..7285)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7459..7479)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7464..7743,1..50)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7644..7743,1..36)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7706..7725
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgcgt
       61 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      121 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      181 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      421 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      481 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      541 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      601 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
      661 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      721 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      781 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      841 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
      901 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
      961 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     1021 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     1081 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     1141 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     1201 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     1261 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     1321 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag
     1381 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     1441 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     1501 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     1561 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     1621 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     1681 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     1741 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     1801 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     1861 gagaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     1921 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     1981 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     2041 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2101 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2161 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     2221 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2281 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2341 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2401 acagcagaga tccagtttgg ttaattaact gcagccccga taaaataaaa gattttattt
     2461 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagctgca
     2521 gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca
     2581 agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt
     2641 tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc
     2701 caagaacaga tggtccccag atatggccca accctcagca gtttcttaag acccatcaga
     2761 tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc
     2821 agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac
     2881 aacccctcac tcggcgcgcc agtcctccga cagactgagt cgcccggggg tctagaagcg
     2941 ctggatccgt ttaaacgcgg ccgcccagca cagtggctcg agccgcgggt taactggcca
     3001 gaattctcga ccttgactgg cggcgcgacc ttgaggcctg cgttcgcctc agttgccccc
     3061 tctgtgcaat ggggagacgc gcctcatcgc ttgacaacgg ccgaagagcc gccgcgcttc
     3121 cgtctcccgc gtgcgcgcgc catgctgccc acccccgttc cgcactgacc ctcccccgtg
     3181 ccccgcgtcc cgtactgccg ccccgccccg agtcccatgc cgcagccacc gcgacggagc
     3241 ccgcaggcgg gaacctgcct ccgcgcgtta gcgcgcacgc gcgcctcatg tgtcgtcccc
     3301 atcagcgccg gcttccgtct ataggccaga tgcactgtca ctctggcgaa gtcgcagacc
     3361 cgattggccg ggacggaggc gcgagaccgg gttgcgggcg gggccgaacg tggtataaaa
     3421 ggggcgggag gccaggctcg tgccgttttg cagacgccac cgccgaggaa aaccgtgtac
     3481 tattagccac gcgtgccacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     3541 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     3601 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     3661 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     3721 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     3781 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     3841 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     3901 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     3961 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     4021 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     4081 cccgcaagcc cggtgcctga gcgatcgcta atcaacctct ggattacaaa atttgtgaaa
     4141 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4201 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4261 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4321 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4381 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4441 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4501 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4561 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4621 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4681 tttgggccgc ctccccgctt aatcgcgtcg agacctagaa aaacatggag caatcacaag
     4741 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     4801 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     4861 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     4921 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
     4981 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
     5041 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
     5101 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
     5161 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
     5221 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5281 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5341 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc
     5401 ccgtttcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5461 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5521 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5581 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5641 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5701 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5761 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5821 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5881 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5941 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6001 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6061 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6121 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6181 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6241 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6301 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6361 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6421 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6481 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6541 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6601 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6661 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6721 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6781 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6841 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6901 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6961 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7021 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7081 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7141 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7201 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     7261 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7321 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7381 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     7441 gccagatata cgcgctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     7501 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     7561 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     7621 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     7681 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     7741 agc
//
