LOCUS       sequence-330869-        9361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human CPSF6 fused to mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9361)
  AUTHORS   Zhong Z, Ning J, Boggs EA, Jang S, Wallace C, Telmer C, Bruchez MP, 
            Ahn J, Engelman AN, Zhang P, Watkins SC, Ambrose Z
  TITLE     Cytoplasmic CPSF6 Regulates HIV-1 Capsid Trafficking and Infection 
            in a Cyclophilin A-Dependent Manner.
  JOURNAL   mBio. 2021 Mar 23;12(2). pii: mBio.03142-20. doi: 
            10.1128/mBio.03142-20.
  PUBMED    33758083
REFERENCE   2  (bases 1 to 9361)
  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: "mBio."; 
            date: "2021-03-23"; volume: "12(2). pii: mBio.03142-20. doi"; pages:
            " 10.1128/mBio.03142-20"
COMMENT     Sequence Label: pSICO-CPSF6-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..9361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1447..1680
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1865..1909
                     /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    2207..2324
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2614..2634
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(2652..2685)
                     /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        2785..3088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3089..3292
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      5137..5146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5143..5853
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      5398..5407
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5862..5895)
                     /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     5922..5938
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6013..6601
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6066..6086)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6484..6495)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6604..6620)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7130..7310
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7372..7960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7452..7471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8131..8922,8923..8991))
                     /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     8754..8773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8992..9096)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9171..9190)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
     1321 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     1381 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     1441 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     1501 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     1561 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     1621 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     1681 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     1741 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     1801 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     1861 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     1921 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     1981 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2041 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2101 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     2161 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2221 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2281 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2341 acagcagaga tccagtttgg ttagtaccgg gcccgctcta gagatccgac gcgccatctc
     2401 taggcccgcg ccggccccct cgcacagact tgtgggagaa gctcggctac tcccctgccc
     2461 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
     2521 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
     2581 gagatacaaa tactaaatta ttattttaaa aaacagcaca aaaggaaact caccctaact
     2641 gtaaagtaat tataacttcg tatagtataa attatacgaa gttataagcc ttggtttttt
     2701 gaattccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     2761 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     2821 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     2881 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     2941 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     3001 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     3061 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     3121 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     3181 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     3241 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     3301 gaaccgtcgg atcccagtgt ggtggtacgg gaattcagga agatggcgga cggcgtggac
     3361 cacatagaca tttacgcgga tgtcggcgaa gagttcaacc aggaagctga atatggtggg
     3421 catgatcaga tagatttgta tgacgatgtc atatctccat ctgcaaataa tggagatgcc
     3481 ccagaagacc gagattacat ggatactctc ccaccaactg ttggtgatga tgtgggtaaa
     3541 ggagcagcac caaatgttgt ctatacatat actggaaaga gaattgcatt atatattgga
     3601 aatctaacat ggtggacaac agatgaagac ttaactgaag cagttcattc tttgggagta
     3661 aatgatattt tggagataaa attttttgaa aatcgggcaa atggccagtc aaaggggttt
     3721 gcccttgttg gtgttggatc tgaagcatct tcaaaaaagt taatggatct gttacctaaa
     3781 agagaacttc atggtcagaa tcctgttgta actccatgca ataaacagtt cctgagtcaa
     3841 tttgaaatgc agtccaggaa aactacacaa tcaggacaaa tgtctgggga aggtaaagct
     3901 ggtcctccag gaggcagttc ccgtgcagca tttccacaag gtggtagagg acggggccgt
     3961 tttccagggg ctgttcctgg tggggacaga tttcctgggc cagcaggacc aggagggcca
     4021 cccccacctt ttccaggaaa tttgatcaag catcttgtta aaggaactcg gcctttgttc
     4081 ctggaaacta ggattccatg gcatatgggg cacagcatag aggaaatacc catttttggc
     4141 ctaaaagctg gacagactcc accacgtcca cccttaggtc ctccaggccc acctggtcca
     4201 ccaggtcctc cacctcctgg tcaggttctg cctcctcctc tagctgggcc tcctaatcga
     4261 ggagatcgcc ctccaccacc agttcttttt cctggacaac cttttgggca gcctccattg
     4321 ggtccacttc ctcctggccc tccacctcca gttccaggct acggcccccc tcctggccca
     4381 ccacctccac aacagggacc acctccacct ccaggcccct ttccacctcg tccacccggt
     4441 ccacttgggc caccccttac actagctcct cctccgcatc ttcctggacc acctccaggt
     4501 gccccaccgc cagctccgca tgtgaaccca gctttctttc ctccaccaac taacagtggc
     4561 atgcctacat cagatagccg aggtccacca ccaacagatc catatgggcg acctccacca
     4621 tatgataggg gtgactatgg cccccctgga agggaaatgg atactgcaag aacgccattg
     4681 agtgaagctg aatttgaaga aatcatgaat agaaataggg caatctcaag cagtgctatt
     4741 tcgagagctg tgtctgatgc cagtgctggt gattatggga gtgctattga gacactggta
     4801 actgcaattt ctttaattaa acaatccaaa gtatctgctg atgatcgttg caaagttctt
     4861 attagttctt tgcaagattg ccttcatgga attgagtcca agtcttatgg ttctggatca
     4921 agacgtgaac gatcaagaga gagggaccat agtagatcac gagaaaagag tcgacgtcat
     4981 aaatcccgta gtagagaccg tcatgacgat tattacagag agagaagcag agaacgagag
     5041 aggcaccggg atcgtgaccg agaccgtgac cgagagcgtg accgagagcg cgaatatcgt
     5101 catcgtgagg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
     5161 gaggataaca tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac
     5221 ggtgtggact ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag
     5281 ttaaacctga agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat
     5341 atcgggtatg gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc
     5401 gccatggtag atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc
     5461 tcccttactg ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag
     5521 gtgaagggga ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg
     5581 gactggtgca ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag
     5641 tggagttaca ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc
     5701 tttgccaagc caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag
     5761 acggagctca agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc
     5821 gatgtgatgg gcatggacga gctgtacaag taagcggccg cataacttcg tatagtataa
     5881 attatacgaa gttataagcc ttgttaacgc gcggtgaccc tcgaggtcga cggtatcgat
     5941 aagctcgctt cacgagatca tgtttaaggg ttccggttcc actaggtaca attcgatatc
     6001 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     6061 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6121 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6181 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6241 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6301 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6361 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     6421 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6481 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6541 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6601 catcgatacc gtcgacctcg atcgagacct agaaaaacat ggagcaatca caagtagcaa
     6661 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     6721 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     6781 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     6841 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
     6901 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
     6961 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
     7021 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
     7081 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
     7141 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7201 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7261 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatgtgagc
     7321 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7381 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7441 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7501 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7561 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7621 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7681 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7741 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7801 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7861 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7921 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7981 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8041 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8101 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8161 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8221 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8281 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8341 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8401 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8461 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8521 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8581 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8641 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8701 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8761 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8821 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8881 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8941 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9061 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9121 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     9181 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     9241 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     9301 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     9361 t
//
