LOCUS       sequence-362810-        7802 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 7802)
  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 7802)
  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 Cullin5
FEATURES             Location/Qualifiers
     source          1..7802
                     /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             4140..4163
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4192..4780
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4245..4265)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4663..4674)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4855..5088
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5207..5225)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5293..5397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5398..5466,5467..6258)
                     /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(5616..5635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6429..7017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6918..6937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7171..7188
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7428..7449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7464..7469,7470..7487,7488..7494)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7502..7518
                     /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     7507..7529
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7526..7542
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7526..7542
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7592..7611
                     /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 cgacgtctaa tctgttaaag aataaaggtt ctcttcagtt tgaagacaaa tgggatttta
     2881 tgcgcccgat tgttttgaag cttttacgcc aggaatctgt tacaaaacag cagtggtttg
     2941 atctgttttc ggatgtgcat gcagtctgtc tttgggatga taaaggccca gcaaaaattc
     3001 atcaggcttt aaaagaagat attcttgagt ttattaagca ggcacaggca cgagtactga
     3061 gccatcaaga tgatacggct ttgctaaaag catatattgt tgaatggcga aagttcttta
     3121 cacaatgtga tattttacca aaaccttttt gtcaactaga gattacttta atgggtaaac
     3181 agggcagcaa taaaaaatca aatgtggaag acagtattgt tcgaaagctt atgcttgata
     3241 catggaatga gtcaatcttt tcaaacataa aaaacagact ccaagatagt gcaatgaagc
     3301 tggtacatgc tgagagattg ggagaagctt ttgattctca gctggttatt ggagtaagag
     3361 aatcctatgt taacctttgt tctaatcctg aggataaact tcaaatttat agggacaatt
     3421 ttgagaaggc atacttggat tcaacagaga gattttatag aacacaagca ccctcgtatt
     3481 tacaacaaaa tggtgtacag aattatatga aatatgcaga tgctaaatta aaagaagaag
     3541 aaaaacgagc actacgttat ttagaaacaa gacgagaatg taactccgtt gaagcactca
     3601 tggaatgctg tgtaaatgcc ctggtgacat catttaaaga gactatctta gctgagtgcc
     3661 aaggcatgat caagagaaat gaaactgaaa aattacattt aatgttttca ttgatggaca
     3721 aagttcctaa tggtatagag ccaatgttga aagacttgga ggaacatatc attagtgctg
     3781 gcctggcaga tatggtagca gctgctgaaa ctattactac tgactctgag aaatacgttg
     3841 agcagttact tacactattt aatagattta gtaaactcgt caaagaagct tttcaagatg
     3901 atccacgatt tcttactgca agagataagg cgtataaagc agttgttaat gatgctacca
     3961 tatttaaact tgaattacct ttgaagcaga agggggtggg attaaaaact cagcctgaat
     4021 caaaatgccc tgagctgctt gccaattact gtgacatgtt gctaagaaaa acaccattaa
     4081 gcaaaaaact aacctctgaa gagattgaag caaagcttaa agaagtgctc ttggtacttg
     4141 actacaagga tgacgatgac aagtagtcga gcatgcatct agatagatcc taatcaacct
     4201 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4261 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4321 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     4381 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4441 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     4501 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     4561 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     4621 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     4681 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4741 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgagatcct ttaagaccaa
     4801 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     4861 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     4921 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     4981 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     5041 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     5101 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     5161 ccgggttaat taaggaaagg gctagatcat tcttgaagac gaaagggcct cgtgatacgc
     5221 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5281 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5341 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5401 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5461 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5521 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5581 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5641 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5701 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5761 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5821 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5881 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5941 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6001 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6241 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6301 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6361 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6541 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6781 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7081 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7141 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7201 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcaagctc
     7261 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     7321 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctccccgtgg
     7381 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7441 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7501 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaatttcaca
     7561 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     7621 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
     7681 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
     7741 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
     7801 gt
//
