LOCUS       sequence-320630-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suppress RTEL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Sviderskiy VO, Blumenberg L, Gorodetsky E, Karakousi TR, Hirsh N, 
            Alvarez SW, Terzi EM, Kaparos E, Whiten GC, Ssebyala S, Tonzi P, Mir
            H, Neel BG, Huang TT, Adams S, Ruggles KV, Possemato R
  TITLE     Hyperactive CDK2 Activity in Basal-like Breast Cancer Imposes a 
            Genome Integrity Liability that Can Be Exploited by Targeting DNA 
            Polymerase epsilon.
  JOURNAL   Mol Cell. 2020 Oct 28. pii: S1097-2765(20)30723-1. doi: 
            10.1016/j.molcel.2020.10.016.
  PUBMED    33152268
REFERENCE   2  (bases 1 to 7084)
  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: "Mol Cell. 
            2020 Oct 28. pii: S1097-2765(20)30723-1. doi: 
            10.1016/j.molcel.2020.10.016."
COMMENT     Sequence Label: pLKO.1P RTEL1_1
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             74..118
                     /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        416..664
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     416..436
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     587..606
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    761..878
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        927..1437
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1356..1374
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1459..2058
                     /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(1459..1478)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1955..1975
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2186..2419
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2491..2612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2528..2547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2582..2601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2652..2787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2807..2826)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2808..2826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2836..2853)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2836..2852)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2845..2867)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2994..3449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3081..3100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3291..3312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3475..3579
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3580..3648,3649..4440)
                     /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(3798..3817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4611..5199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5100..5119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5353..5370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5487..5508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5523..5528,5529..5546,5547..5553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5561..5577
                     /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     5566..5588
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5585..5601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5585..5601
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5620..5640
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5622..5640
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5668..5894
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5895..6075
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6122..6247
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6740..6973
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
       61 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
      121 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
      181 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
      241 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
      301 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
      361 cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg
      421 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
      481 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
      541 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
      601 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
      661 caccggccaa caggagtaca tgaccaactc gagttggtca tgtactcctg ttggtttttg
      721 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
      781 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      841 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      901 gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca aggcagccct
      961 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac
     1021 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc
     1081 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg
     1141 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac
     1201 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc
     1261 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt
     1321 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg
     1381 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg
     1441 gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     1501 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     1561 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     1621 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     1681 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     1741 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     1801 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     1861 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     1921 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     1981 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     2041 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
     2101 cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     2161 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     2221 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     2281 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     2341 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     2401 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     2461 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     2521 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2581 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     2641 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2701 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2761 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     2821 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2881 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2941 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     3001 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3061 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3121 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3181 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3241 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3301 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3361 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3421 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     3481 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3541 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3601 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3661 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3721 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3781 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3841 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3901 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3961 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4021 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4081 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4141 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4201 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4261 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4321 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4381 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4441 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4501 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4561 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4621 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4681 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4741 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4801 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4861 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4921 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4981 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5041 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5101 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5161 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5221 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5281 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5341 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     5401 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5461 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     5521 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     5581 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     5641 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     5701 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     5761 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     5821 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     5881 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5941 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6001 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6061 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     6121 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     6181 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     6241 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     6301 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     6361 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     6421 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     6481 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     6541 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     6601 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     6661 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     6721 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     6781 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     6841 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     6901 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     6961 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     7021 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     7081 tgga
//
