LOCUS       sequence-271840-        7301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid expressing sgRNA against HJURP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7301)
  AUTHORS   Swartz SZ, McKay LS, Su KC, Bury L, Padeganeh A, Maddox PS, Knouse 
            KA, Cheeseman IM
  TITLE     Quiescent Cells Actively Replenish CENP-A Nucleosomes to Maintain 
            Centromere Identity and Proliferative Potential.
  JOURNAL   Dev Cell. 2019 Oct 7;51(1):35-48.e7. doi: 
            10.1016/j.devcel.2019.07.016. Epub 2019 Aug 15.
  PUBMED    31422918
REFERENCE   2  (bases 1 to 7301)
  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: 
            "10.1016/j.devcel.2019.07.016"; journalName: "Dev Cell"; date: 
            "2019-10-7- 7"; volume: "51"; issue: "1"; pages: "35-48.e7"
COMMENT     Sequence Label: pZS160
FEATURES             Location/Qualifiers
     source          1..7301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    177..294
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        363..574
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             600..1199
                     /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(600..619)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1096..1116
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1215..1803
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1268..1288)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1686..1697)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1875..2108
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2186..2307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2223..2242)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2277..2296
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2347..2482
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2502..2521)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2503..2521)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2531..2548)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2531..2547)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2540..2562)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2689..3144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2776..2795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2986..3007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3170..3274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3275..3343,3344..4135)
                     /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     complement(3493..3512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4306..4894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4795..4814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5048..5065
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5182..5203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5218..5223,5224..5241,5242..5248)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5256..5272
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5261..5283
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5280..5296
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5280..5296
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5315..5335
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5317..5335
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5363..5589
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5590..5770
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5817..5942
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6435..6668
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6853..6897
                     /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"
     promoter        join(7075..7301,1..22)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7075..7095
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7246..7265
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 cttgtggaaa ggacgaaaca ccgcccttcg aggacacccc gggtttaaga gctatgctgg
       61 aaacagcata gcaagtttaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
      121 gtcggtgctt tttttgaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt
      181 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
      241 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
      301 tattacaggg acagcagaga tccactttgg cgccggctcg agtggctccg gtgcccgtca
      361 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      421 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
      481 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
      541 tctttttcgc aacgggtttg ccgccagaac acaggtgtcg tgacgcggga tccgccacca
      601 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
      661 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
      721 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
      781 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
      841 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
      901 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
      961 ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag ggtctgggca
     1021 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     1081 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     1141 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa
     1201 cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1261 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1321 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     1381 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     1441 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     1501 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1561 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     1621 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     1681 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     1741 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     1801 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     1861 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     1921 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     1981 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     2041 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     2101 ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     2161 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     2221 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2281 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     2341 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2401 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2461 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     2521 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     2581 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     2641 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     2701 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2761 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2821 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2881 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2941 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3001 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3061 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3121 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3181 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3241 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3301 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3361 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3421 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3481 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3541 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3601 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3661 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3721 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3781 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3841 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3901 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3961 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4021 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4081 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4141 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4201 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4321 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4381 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4441 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4501 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4561 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4621 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4681 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4741 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4801 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4861 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4921 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4981 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5041 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5101 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5161 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5221 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5281 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     5341 aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg
     5401 gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg
     5461 gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg
     5521 attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga
     5581 tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5641 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     5701 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     5761 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     5821 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     5881 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     5941 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     6001 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     6061 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     6121 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     6181 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     6241 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     6301 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     6361 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     6421 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     6481 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     6541 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     6601 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     6661 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     6721 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     6781 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     6841 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     6901 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     6961 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     7021 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cagagagggc
     7081 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
     7141 agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
     7201 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
     7261 accgtaactt gaaagtattt cgatttcttg gctttatata t
//
