LOCUS       sequence-440849-        8759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector that expresses sgRNA resistant DERA in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8759)
  AUTHORS   Zhao Y, Tabet D, Rubio Contreras D, Lao L, Kousholt AN, Weile J, 
            Melo H, Hoeg L, Feng S, Cote AG, Lin ZY, Setiaputra D, Jonkers J, 
            Gingras AC, Gomez Herreros F, Roth FP, Durocher D
  TITLE     Genome-scale mapping of DNA damage suppressors through phenotypic 
            CRISPR-Cas9 screens.
  JOURNAL   Mol Cell. 2023 Aug 3;83(15):2792-2809.e9. doi: 
            10.1016/j.molcel.2023.06.025. Epub 2023 Jul 20.
  PUBMED    37478847
REFERENCE   2  (bases 1 to 8759)
  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.2023.06.025"; journalName: "Mol Cell"; date: 
            "2023-08-3- 3"; volume: "83"; issue: "15"; pages: "2792-2809.e9"
COMMENT     Sequence Label: pHIV(T9)-DERA-sgresis
FEATURES             Location/Qualifiers
     source          1..8759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             588..641
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     LTR             1370..1550
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(1612..2200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1692..1711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2371..3162,3163..3231))
                     /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     2994..3013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3232..3336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3411..3430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3602..3981
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3983..4181
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4136..4156
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4199..4379
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4426..4551
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5048..5281
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5466..5510
                     /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    5808..5925
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5988..7166
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6219..7157
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7114..7134
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(8181..8755,8756..8758,8759,1..8)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8348..8365)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8575..8594
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 ccacaaccat gggcaccacc ctggacgaca cggcttaccg ataccgcacc agtgtccctg
       61 gggacgccga ggccatcgag gcactggatg ggtccttcac caccgacacc gtcttccgcg
      121 tcaccgccac cggggacggc ttcaccctgc gggaggtgcc ggtggacccg cccctgacca
      181 aggtgttccc cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgacccgg
      241 actcccggac gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct
      301 cgtactccgg ctggaaccgc cggctgaccg tcgaggacat tgaggtagca cccgagcaca
      361 ggggacacgg ggtgggacgg gcattgatgg gactggctac cgagttcgcc agagaacggg
      421 gagctggtca tctttggctc gaagtcacca acgtcaacgc accggcgatc cacgcttacc
      481 ggcggatggg gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg
      541 acggcgagca ggcgctctac atgtccatgc cctgccccgg cagtggcgag ggcagaggaa
      601 gtctgctaac atgcggtgac gtcgaggaga atcctggccc aatgaagcgc ttcctcttcc
      661 tcctactcac catcagcctc ctggttatgg tacagataca aactggactc tcaggacaaa
      721 acgacaccag ccaaaccagc agcccctcag catccagcaa cataagcgga ggcattttcc
      781 ttttcttcgt ggctaatgcc ataatccacc tcttctgctt cagttgaggc gcgccaaatt
      841 tatcgatacc gtcgacttcg atcgagacct agaaaaacat ggagcaatca caagtagcaa
      901 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
      961 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     1021 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     1081 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
     1141 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
     1201 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
     1261 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
     1321 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
     1381 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     1441 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     1501 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatgtgagc
     1561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3361 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     3421 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     3481 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     3541 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     3601 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     3661 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     3721 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     3781 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     3841 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     3901 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     3961 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     4021 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     4081 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     4141 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
     4201 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     4261 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     4321 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     4381 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     4441 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     4501 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     4561 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     4621 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     4681 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     4741 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     4801 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     4861 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag
     4921 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     4981 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     5041 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     5101 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     5161 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     5221 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     5281 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     5341 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     5401 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     5461 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     5521 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     5581 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     5641 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     5701 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     5761 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     5821 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     5881 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     5941 gacagcagag atccagtttg gttagtaccg ggcccgctct agcgtgaggc tccggtgccc
     6001 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     6061 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     6121 ggctccgcct ttttcccgag ggtgggggag aaccgtaccc ccgtgcagta gtcgccgtga
     6181 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     6241 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
     6301 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     6361 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     6421 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     6481 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     6541 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     6601 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     6661 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     6721 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     6781 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     6841 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     6901 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     6961 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     7021 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     7081 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     7141 tttttcttcc atttcaggtg tcgtgagcgg ccgcaccatg tccgcgcaca atcggggcac
     7201 cgagctcgac cttagctgga tctccaaaat acaagtgaat cacccggcag ttctgaggcg
     7261 tgcggaacaa atccaggctc gcagaaccgt gaaaaaggag tggcaggctg cttggctcct
     7321 gaaagctgtt acctttatag atcttactac actttcaggt gatgatacat cttcaaacat
     7381 tcaaaggctc tgttataaag ccaaataccc aatccgggaa gatctcttaa aagctttaaa
     7441 tatgcatgat aaaggcatta ctacagccgc cgtttgtgtt tatcccgccc gggtgtgtga
     7501 tgctgtaaaa gcactcaagg ctgcaggctg taatatccct gtggcatcag tggccgctgg
     7561 atttccagct ggacagactc atttgaagac acgattagaa gagatcagat tggctgtgga
     7621 agatggagct acagaaatcg acgtggtaat taacagaagc ttggtgctga caggccagtg
     7681 ggaagccctg tatgatgaga ttcgtcagtt tcgcaaggcc tgtggggagg ctcatcttaa
     7741 aactatatta gcgacaggag aacttggaac tcttactaat gtctataaag ccagtatgat
     7801 agcaatgatg gcaggatcag attttattaa gacctctact ggaaaagaaa cagtaaatgc
     7861 caccttcccg gtagctatag taatgctgcg ggccattaga gatttcttct ggaaaactgg
     7921 aaacaagata gggtttaaac cagcaggagg catccgcagt gcaaaggatt cccttgcttg
     7981 gctctctctt gtaaaggagg agcttggaga tgagtggctg aagccagaac tctttcgaat
     8041 aggtgccagt actctgctct cggacattga gaggcagatt taccatcatg tgactggaag
     8101 atatgcagct tatcatgatc ttccaatgtc ttgagaattc atcgacgtta actattctag
     8161 agtacccggg ctaggatccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     8221 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     8281 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     8341 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     8401 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     8461 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     8521 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     8581 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     8641 tctgatctgg ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     8701 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatgg
//
