LOCUS       sequence-333139-        7758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre recombinase and an sgRNA targeting Zmat3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7758)
  AUTHORS   Bieging-Rolett KT, Kaiser AM, Morgens DW, Boutelle AM, Seoane JA, 
            Van Nostrand EL, Zhu C, Houlihan SL, Mello SS, Yee BA, McClendon J, 
            Pierce SE, Winters IP, Wang M, Connolly AJ, Lowe SW, Curtis C, Yeo 
            GW, Winslow MM, Bassik MC, Attardi LD
  TITLE     Zmat3 Is a Key Splicing Regulator in the p53 Tumor Suppression 
            Program.
  JOURNAL   Mol Cell. 2020 Nov 5;80(3):452-469.e9. doi: 
            10.1016/j.molcel.2020.10.022.
  PUBMED    33157015
REFERENCE   2  (bases 1 to 7758)
  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.molcel.2020.10"; journalName: "Mol Cell"; date: 
            "2020-11-5- 5"; volume: "80"; issue: "3"; pages: "452-469.e9"
COMMENT     Sequence Label: Lenti-sgZmat3.51.5/Cre
FEATURES             Location/Qualifiers
     source          1..7758
                     /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"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     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"
     promoter        2406..2654
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2406..2426
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2577..2596
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2674..2749
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2802..3301
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2837..2860)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3234..3253
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3305..3325
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3326..4354
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    4410..4998
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4463..4483)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4881..4892)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5001..5017)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5002..5018)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5527..5707
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5769..6357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5849..5868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6528..7319,7320..7388))
                     /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     7151..7170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7389..7493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7568..7587)
                     /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 gccctgcagg aactgtttaa
     2401 actgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     2461 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     2521 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     2581 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     2641 aaggaccaaa caccggctta cacagctcct ccagttttag agctagaaat agcaagttaa
     2701 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttg
     2761 gcgcgcctat ggcttaatta acatggcggc cgcattctac cgggtagggg aggcgctttt
     2821 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     2881 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     2941 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     3001 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     3061 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     3121 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     3181 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     3241 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggacccgg
     3301 gatgcccaag aagaagagga aggtgtccaa tttactgacc gtacaccaaa atttgcctgc
     3361 attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga
     3421 tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc
     3481 ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag atgttcgcga
     3541 ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa actatccagc aacatttggg
     3601 ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt
     3661 ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca
     3721 ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg aaaatagcga
     3781 tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca ccctgttacg
     3841 tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg gtgggagaat
     3901 gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag agaaggcact
     3961 tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg tagctgatga
     4021 tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc catctgccac
     4081 cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc gattgattta
     4141 cggcgctaag gatgactctg gtcagagata cctggcctgg tctggacaca gtgcccgtgt
     4201 cggagccgcg cgagatatgg cccgcgctgg agtttcaata ccggagatca tgcaagctgg
     4261 tggctggacc aatgtaaata ttgtcatgaa ctatatccgt aacctggata gtgaaacagg
     4321 ggcaatggtg cgcctgctgg aagatggcga ttagccatgt ttaagggttc cggttccact
     4381 aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     4441 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4501 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4561 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4621 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4681 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4741 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4801 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4861 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4921 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4981 tttgggccgc ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga
     5041 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
     5101 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
     5161 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     5221 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     5281 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg
     5341 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac
     5401 acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta
     5461 gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac
     5521 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5581 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5641 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     5701 tctagcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5761 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5821 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5881 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5941 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6001 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6061 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7501 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     7561 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     7621 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7681 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     7741 gggccagata tacgcgtt
//
