LOCUS       sequence-208230-        8691 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 8691)
  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 8691)
  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..8691
                     /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    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2854..2874
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(2892..2925)
                     /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        3025..3328
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3329..3532
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      4696..4705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4702..4704,4705..4707,4708..5421)
                     /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(4702..4704,4705..4707,4708..5418)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4747..4768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5008..5027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5355..5376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5429..5462)
                     /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     5489..5505
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5580..6168
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5633..5653)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6051..6062)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6171..6187)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6697..6877
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6939..7527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7019..7038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7698..8489,8490..8558))
                     /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     8321..8340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8559..8663)
                     /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 gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccggg cccgctctag agatccgacg ccgccatctc
     2641 taggcccgcg ccggccccct cgcacagact tgtgggagaa gctcggctac tcccctgccc
     2701 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
     2761 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
     2821 gagatacaaa tactaaatta ttattttaaa aaacagcaca aaaggaaact caccctaact
     2881 gtaaagtaat tataacttcg tatagtataa attatacgaa gttataagcc ttggtttttt
     2941 gaattccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     3001 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     3061 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3121 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     3181 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     3241 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     3301 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     3361 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     3421 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     3481 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     3541 gaaccgtcgg atcccagtgt ggtggtacgg gaattcagga agatggcgga cggcgtggac
     3601 cacatagaca tttacgcgga tgtcggcgaa gagttcaacc aggaagctga atatggtggg
     3661 catgatcaga tagatttgta tgacgatgtc atatctccat ctgcaaataa tggagatgcc
     3721 ccagaagacc gagattacat ggatactctc ccaccaactg ttggtgatga tgtgggtaaa
     3781 ggagcagcac caaatgttgt ctatacatat actggaaaga gaattgcatt atatattgga
     3841 aatctaacat ggtggacaac agatgaagac ttaactgaag cagttcattc tttgggagta
     3901 aatgatattt tggagataaa attttttgaa aatcgggcaa atggccagtc aaaggggttt
     3961 gcccttgttg gtgttggatc tgaagcatct tcaaaaaagt taatggatct gttacctaaa
     4021 agagaacttc atggtcagaa tcctgttgta actccatgca ataaacagtt cctgagtcaa
     4081 tttgaaatgc agtccaggaa aactacacaa tcaggacaaa tgtctgggga aggtaaagct
     4141 ggtcctccag gaggcagttc ccgtgcagca tttccacaag gtggtagagg acggggccgt
     4201 tttccagggg ctgttcctgg tggggacaga tttcctgggc cagcaggacc aggagggcca
     4261 cccccacctt ttccaggaaa tttgatcaag catcttgtta aaggaactcg gcctttgttc
     4321 ctggaaacta ggattccatg gcatatgggg cacagcatag aggaaatacc catttttggc
     4381 ctaaaagctg gacagactcc accacgtcca cccttaggtc ctccaggccc acctggtcca
     4441 ccaggtcctc cacctcctgg tcaggttctg cctcctcctc tagctgggcc tcctaatcga
     4501 ggagatcgcc ctccaccacc agttcttttt cctggacaac cttttgggca gcctccattg
     4561 ggtccacttc ctcctggccc tccacctcca gttccaggct acggcccccc tcctggccca
     4621 ccacctccac aacagggacc acctccacct ccaggcgagg taccgcgggc ccgggatccg
     4681 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     4741 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4801 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4861 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     4921 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4981 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     5041 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5101 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5161 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     5221 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     5281 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     5341 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     5401 atggacgagc tgtacaagta gcggccgcat aacttcgtat agtataaatt atacgaagtt
     5461 ataagccttg ttaacgcgcg gtgaccctcg aggtcgacgg tatcgataag ctcgcttcac
     5521 gagatcatgt ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata
     5581 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5641 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5701 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5761 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5821 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5881 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5941 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     6001 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6061 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6121 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     6181 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     6241 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     6301 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6361 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     6421 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     6481 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     6541 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     6601 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     6661 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     6721 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6781 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6841 ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa
     6901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7201 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7321 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7501 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7561 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7621 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7681 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7741 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7801 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7861 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7921 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7981 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8041 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8101 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8161 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8221 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8281 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8341 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8401 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8461 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8521 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8581 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8641 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
