LOCUS       sequence-213756-        8688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a truncated form of CPSF6 (CPSF6-358 originally described 
            in Lee et al. Cell Host Microbe 2010) fused with eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8688)
  AUTHORS   Ning J, Zhong Z, Fischer DK, Harris G, Watkins SC, Ambrose Z, Zhang 
            P
  TITLE     Truncated CPSF6 forms higher order complexes that bind and disrupt 
            HIV-1 capsid.
  JOURNAL   J Virol. 2018 Apr 11. pii: JVI.00368-18. doi: 10.1128/JVI.00368-18.
  PUBMED    29643241
REFERENCE   2  (bases 1 to 8688)
  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"; journalName: "J Virol. 
            2018 Apr 11. pii: JVI.00368-18. doi: 10.1128/JVI.00368-18."
COMMENT     Sequence Label: pSICO-CPSF6-358-eGFP
FEATURES             Location/Qualifiers
     source          1..8688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2851..2871
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(2889..2922)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     enhancer        3022..3325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3326..3529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      4693..4702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4699..4701,4702..4704,4705..5418)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4699..4701,4702..4704,4705..5415)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4744..4765)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5005..5024)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5352..5373
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5426..5459)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     primer_bind     5486..5502
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5577..6165
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5630..5650)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6048..6059)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6168..6184)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6694..6874
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6936..7524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7016..7035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7695..8486,8487..8555))
                     /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     8318..8337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8556..8660)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cgctctagag atccgacgcg ccatctctag
     2641 gcccgcgccg gccccctcgc acagacttgt gggagaagct cggctactcc cctgccccgg
     2701 ttaatttgca tataatattt cctagtaact atagaggctt aatgtgcgat aaaagacaga
     2761 taatctgttc tttttaatac tagctacatt ttacatgata ggcttggatt tctataagag
     2821 atacaaatac taaattatta ttttaaaaaa cagcacaaaa ggaaactcac cctaactgta
     2881 aagtaattat aacttcgtat agtataaatt atacgaagtt ataagccttg gttttttgaa
     2941 ttccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     3001 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     3061 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     3121 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     3181 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     3241 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     3301 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     3361 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3421 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     3481 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     3541 ccgtcggatc ccagtgtggt ggtacgggaa ttcaggaaga tggcggacgg cgtggaccac
     3601 atagacattt acgcggatgt cggcgaagag ttcaaccagg aagctgaata tggtgggcat
     3661 gatcagatag atttgtatga cgatgtcata tctccatctg caaataatgg agatgcccca
     3721 gaagaccgag attacatgga tactctccca ccaactgttg gtgatgatgt gggtaaagga
     3781 gcagcaccaa atgttgtcta tacatatact ggaaagagaa ttgcattata tattggaaat
     3841 ctaacatggt ggacaacaga tgaagactta actgaagcag ttcattcttt gggagtaaat
     3901 gatattttgg agataaaatt ttttgaaaat cgggcaaatg gccagtcaaa ggggtttgcc
     3961 cttgttggtg ttggatctga agcatcttca aaaaagttaa tggatctgtt acctaaaaga
     4021 gaacttcatg gtcagaatcc tgttgtaact ccatgcaata aacagttcct gagtcaattt
     4081 gaaatgcagt ccaggaaaac tacacaatca ggacaaatgt ctggggaagg taaagctggt
     4141 cctccaggag gcagttcccg tgcagcattt ccacaaggtg gtagaggacg gggccgtttt
     4201 ccaggggctg ttcctggtgg ggacagattt cctgggccag caggaccagg agggccaccc
     4261 ccaccttttc caggaaattt gatcaagcat cttgttaaag gaactcggcc tttgttcctg
     4321 gaaactagga ttccatggca tatggggcac agcatagagg aaatacccat ttttggccta
     4381 aaagctggac agactccacc acgtccaccc ttaggtcctc caggcccacc tggtccacca
     4441 ggtcctccac ctcctggtca ggttctgcct cctcctctag ctgggcctcc taatcgagga
     4501 gatcgccctc caccaccagt tctttttcct ggacaacctt ttgggcagcc tccattgggt
     4561 ccacttcctc ctggccctcc acctccagtt ccaggctacg gcccccctcc tggcccacca
     4621 cctccacaac agggaccacc tccacctcca ggcgaggtac cgcgggcccg ggatccgcta
     4681 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     4741 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     4801 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4861 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     4921 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     4981 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     5041 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     5101 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     5161 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     5221 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     5281 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     5341 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     5401 gacgagctgt acaagtagcg gccgcataac ttcgtatagt ataaattata cgaagttata
     5461 agccttgtta acgcgcggtg accctcgagg tcgacggtat cgataagctc gcttcacgag
     5521 atcatgttta agggttccgg ttccactagg tacaattcga tatcaagctt atcgataatc
     5581 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5641 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5701 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5761 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5821 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5881 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5941 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     6001 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     6061 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     6121 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     6181 ctcgatcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     6241 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     6301 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6361 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     6421 tggatctacc acacacaagg ctacttccct gattggcaga actacacacc agggccaggg
     6481 atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag
     6541 gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg
     6601 atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat
     6661 cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga
     6721 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6781 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6841 agaccctttt agtcagtgtg gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag
     6901 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6961 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7021 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7081 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7141 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7201 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7261 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7321 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     7381 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7441 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7501 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7561 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7621 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7681 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7741 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7801 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7861 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7921 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7981 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8041 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8101 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8161 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8221 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8281 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8341 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8401 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8461 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8521 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8581 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8641 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
