LOCUS       sequence-362819-        7801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7801)
  AUTHORS   Emanuele MJ, Elia AE, Xu Q, Thoma CR, Izhar L, Leng Y, Guo A, Chen 
            YN, Rush J, Hsu PW, Yen HC, Elledge SJ
  TITLE     Global identification of modular cullin-RING ligase substrates.
  JOURNAL   Cell. 2011 Oct 14;147(2):459-74. doi: 10.1016/j.cell.2011.09.019. 
            Epub 2011 Sep 29.
  PUBMED    21963094
REFERENCE   2  (bases 1 to 7801)
  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.cell.2011.09.019"; journalName: "Cell"; date: 
            "2011-10-14- 14"; volume: "147"; issue: "2"; pages: "459-74"
COMMENT     Sequence Label: pHAGE DN Cullin4A
FEATURES             Location/Qualifiers
     source          1..7801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2227..2530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2531..2734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2685..2705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2731..2755
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4137..4160
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4191..4779
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4244..4264)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4662..4673)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4854..5087
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5206..5224)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5292..5396
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5397..5465,5466..6257)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5615..5634)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6428..7016
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6917..6936
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7170..7187
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7427..7448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7463..7468,7469..7486,7487..7493)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7501..7517
                     /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     7506..7528
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7525..7541
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7525..7541
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7591..7610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccga tccaccatgg
     2821 cggacgaggc cccgcggaag ggcagcttct cggcgctcgt gggccgcacc aacggcctca
     2881 ccaagcccgc ggccctggcc gccgcgcccg ccaagccggg gggcgcgggc ggctccaaga
     2941 agctggtcat caagaacttc cgagacagac ctcggctgcc cgacaactac acgcaggaca
     3001 cgtggcggaa gctgcacgag gcggtgcggg ccgtgcagag cagcacctcc atcaggtaca
     3061 acctcgagga gctctaccag gctgtggaaa atctctgttc tcacaaagtc tccccaatgc
     3121 tctacaagca actgcgtcag gcctgtgaag accacgtcca ggcacagatc cttccgttta
     3181 gagaagactc actagatagt gttttatttt taaagaagat taacacgtgc tggcaggacc
     3241 actgcagaca aatgatcatg atcagaagca tcttcctgtt cttggaccgc acctatgtgc
     3301 tgcagaactc cacgctgccc tccatctggg atatgggatt agaactgttt agaacccata
     3361 ttattagtga taaaatggtt cagagtaaaa ccattgatgg aatcctactg ctgatcgagc
     3421 gcgagaggag cggcgaggcc gtggaccgga gcctgttgcg gagcctcctg ggcatgctgt
     3481 ctgacctgca ggtgtataaa gattcatttg aactgaaatt tttggaagag actaattgct
     3541 tatatgctgc cgaaggccaa aggttaatgc aggaaagaga ggtcccagaa tatcttaacc
     3601 atgtaagtaa acgcttagag gaagagggag acagagtaat cacttacttg gaccacagca
     3661 cacagaaacc actgattgct tgtgtggaga aacagctatt aggagaacat ttaacagcaa
     3721 ttctgcagaa agggctcgac cacttactgg atgagaacag agtgccggac ctcgcacaga
     3781 tgtaccagct gttcagccgg gtgaggggcg ggcagcaggc gctgctgcag cactggagcg
     3841 agtacatcaa gacttttgga acagcgatcg taatcaatcc tgagaaagat aaagacatgg
     3901 tccaagacct gttggacttc aaggacaagg tggaccacgt gatcgaggtc tgcttccaga
     3961 agaatgagcg gttcgtcaac ctgatgaagg agtcctttga gacgttcatc aacaagagac
     4021 ccaacaagcc tgcagaactg atcgcaaagc atgtggattc aaagttaaga gcaggcaaca
     4081 aagaagccac agacgaggag ctggagcgga cgttggacaa gatcatgatc ctgttcgact
     4141 acaaggatga cgatgacaag tagtcgagca tgcatctagc gatagatcct aatcaacctc
     4201 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4261 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4321 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4381 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4441 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4501 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4561 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     4621 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4681 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4741 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tgagatcctt taagaccaat
     4801 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     4861 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     4921 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     4981 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5041 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5101 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc
     5161 cgggttaatt aaggaaaggg ctagatcatt cttgaagacg aaagggcctc gtgatacgcc
     5221 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5281 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5341 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5401 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5461 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5521 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5581 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     5641 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5701 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5761 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5821 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5881 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5941 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6001 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6061 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6121 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6181 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6241 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6301 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6361 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6421 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6481 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6541 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6601 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6661 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6721 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6781 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6841 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6901 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6961 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7021 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7081 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7141 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     7201 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agcaagctca
     7261 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     7321 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccgtggc
     7381 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7441 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7501 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttcacaa
     7561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7621 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     7681 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
     7741 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     7801 t
//
