LOCUS       sequence-362807-        8263 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 8263)
  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 8263)
  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 Cullin4B
FEATURES             Location/Qualifiers
     source          1..8263
                     /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             4599..4622
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4653..5241
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4706..4726)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5124..5135)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5316..5549
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5668..5686)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5754..5858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5859..5927,5928..6719)
                     /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(6077..6096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6890..7478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7379..7398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7632..7649
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7889..7910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7925..7930,7931..7948,7949..7955)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7963..7979
                     /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     7968..7990
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7987..8003
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7987..8003
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8053..8072
                     /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 tccaccatga
     2821 tgtcacagtc atctggatca ggagatggga atgatgatga ggctactacc tctaaagacg
     2881 gtggtttttc ttcccccagt ccctcagctg ctgctgctgc tcaggaggtc agatctgcca
     2941 ctgatggtaa taccagcacc actccgccca cctctgccaa gaagagaaag ttaaacagca
     3001 gcagcagtag cagcagtaac agtagtaacg agagagaaga ctttgattcc acctcttcct
     3061 cctcttccac tcctccttta caacccaggg attcggcatc cccttcaacc tcgtccttct
     3121 gcctgggggt ttcagtggct gcttccagcc acgtaccgat acagaagaag ctgcgttttg
     3181 aagacaccct ggagtttgta gggtttgatg cgaagatggc tgaggaatcc tcctcctcct
     3241 cctcctcatc ttcaccaact gctgcaacat ctcagcagca gcaacttaaa aataagagta
     3301 tattaatctc ttctgtggct tcggtgcatc atgcaaacgg cctagccaaa tcttctacca
     3361 ccgtctctag ctttgctaac agcaaacctg gctctgctaa gaagttagtg atcaagaact
     3421 ttaaagataa gcctaaatta ccagaaaact acacagatga aacctggcaa aaactgaaag
     3481 aagcagtgga agctattcag aatagtactt caattaagta caatttagaa gaactctacc
     3541 aggctgtaga aaatctctgt tcttacaaga tttctgcaaa cttgtacaaa cagctgagac
     3601 agatctgcga agatcacatc aaagcacaga ttcatcaatt cagagaggat tcattggata
     3661 gcgttctttt tttaaagaag attgatagat gctggcaaaa ccattgcaga caaatgatca
     3721 tgatcaggag catttttttg tttctggata gaacttacgt tcttcagaat tcaatgctac
     3781 cctccatttg ggacatggga ctggagttat ttagggctca tattataagt gatcagaaag
     3841 tgcagaataa gacaattgat ggcattcttc tcttgattga gagggaaagg aatggtgaag
     3901 caattgatag aagtttactt cgaagccttt taagcatgct gtctgatttg caaatttatc
     3961 aagattcttt tgaacaacga tttttggaag aaactaaccg gctctatgca gctgaaggcc
     4021 aaaaattaat gcaagaaaga gaggttcctg aatatctaca tcatgttaac aaacgtctag
     4081 aagaagaagc agacagactt attacttact tagatcagac cacccagaag tcattaattg
     4141 ctactgtaga aaaacaactt ctaggtgaac acttaacagc aattcttcag aaaggtttaa
     4201 ataacctcct tgatgaaaac cgaattcaag atttgtctct tctgtatcag ctcttcagta
     4261 gagttcgagg tggagttcag gttcttttgc agcagtggat cgaatatatc aaggcatttg
     4321 gcagcactat tgtaattaat cctgaaaaag ataaaaccat ggttcaagaa ttgctggatt
     4381 ttaaagataa ggttgaccat ataattgata tctgctttct gaagaatgag aaatttatca
     4441 atgccatgaa agaagcattt gaaacgttca ttaacaaaag accaaataaa ccagctgaac
     4501 ttatagctaa gtatgtagat tcaaaacttc gtgcaggcaa caaagaagct acagatgaag
     4561 aacttgagaa aatgttggat aaaattatga tcatatttga ctacaaggat gacgatgaca
     4621 agtagtcgag catgcatcta gcgatagatc ctaatcaacc tctggattac aaaatttgtg
     4681 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     4741 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     4801 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4861 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4921 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     4981 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5041 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5101 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5161 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     5221 ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca aggcagctgt
     5281 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     5341 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     5401 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     5461 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     5521 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     5581 tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa ttaaggaaag
     5641 ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5701 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5761 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5821 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5881 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5941 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6001 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6061 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     6121 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6181 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6241 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6301 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6361 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6421 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6481 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6541 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6601 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6661 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6721 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6781 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6841 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6901 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6961 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7021 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7081 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7141 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7201 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7261 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7321 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7381 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7441 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7501 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7561 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7621 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7681 ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac taattttttt
     7741 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     7801 cttttttgga ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg tttcccgact
     7861 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7921 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7981 ttcacacagg aaacagctat gacatgatta cgaatttcac aaataaagca tttttttcac
     8041 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
     8101 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
     8161 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
     8221 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
