LOCUS       sequence-362813-        7747 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 7747)
  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 7747)
  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 Cullin2
FEATURES             Location/Qualifiers
     source          1..7747
                     /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             4083..4106
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4137..4725
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4190..4210)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4608..4619)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4800..5033
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5152..5170)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5238..5342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5343..5411,5412..6203)
                     /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(5561..5580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6374..6962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6863..6882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7116..7133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7373..7394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7409..7414,7415..7432,7433..7439)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7447..7463
                     /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     7452..7474
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7471..7487
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7471..7487
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7537..7556
                     /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 tccaccatgt
     2821 ctttgaaacc aagagtagta gattttgatg aaacatggaa caaacttttg acgacaataa
     2881 aagccgtggt catgttggaa tacgtcgaaa gagcaacatg gaatgaccgt ttctcagata
     2941 tctatgcttt atgtgtggcc tatcctgaac cccttggaga aagactttat acagaaacta
     3001 agattttttt ggaaaatcat gttcggcatt tgcataagag agttttggag tcagaagaac
     3061 aagtacttgt tatgtatcat aggtactggg aagaatacag caagggtgca gactatatgg
     3121 actgcttata taggtatctc aacacccagt ttattaaaaa gaataaatta acagaagcgg
     3181 accttcagta tggctatggt ggtgtagata tgaatgaacc acttatggaa ataggagagc
     3241 tagcattgga tatgtggagg aaattgatgg ttgaaccact tcaggccatc cttatccgaa
     3301 tgctgctccg agaaatcaaa aatgatcgtg gtggagaaga cccaaaccag aaagtaatcc
     3361 atggggttat taactccttt gttcatgttg aacagtataa gaaaaaattc cccttaaagt
     3421 tttatcagga aatttttgag tctccctttc tgactgaaac aggagagtat tacaaacaag
     3481 aagcttcaaa tttattacaa gaatcaaact gctcacagta tatggaaaag gttctaggta
     3541 gattaaaaga tgaagaaatt cgatgtcgaa aatacctaca tccaagttcg tatactaagg
     3601 tgattcatga atgtcaacaa cgaatggtag cagaccactt acagttttta catgcagaat
     3661 gtcataatat aattcgacaa gagaaaaaaa atgacatggc aaatatgtac gtcttactcc
     3721 gtgctgtgtc cactggttta cctcatatga ttcaggagct gcaaaaccac atccatgatg
     3781 agggccttcg agcaaccagc aaccttactc aggaaaacat gccaacacta tttgtggagt
     3841 cagttttgga agtgcatggt aaatttgttc agcttatcaa cactgttttg aatggtgatc
     3901 agcattttat gagtgcgttg gataaggccc ttacgtcagt tgtaaattac agagaaccta
     3961 agtctgtttg caaagcacct gaactgcttg ctaagtactg tgacaactta ctgaagaagt
     4021 cagcgaaagg gatgacagag aatgaagtgg aagacaggct cacgagcttc atcacagtgt
     4081 tcgactacaa ggatgacgat gacaagtagt cgagcatgca tctagcgata gatcctaatc
     4141 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4201 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4261 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4321 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4381 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4441 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4501 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     4561 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4621 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4681 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgag atcctttaag
     4741 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     4801 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     4861 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     4921 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     4981 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     5041 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     5101 gaggcccggg ttaattaagg aaagggctag atcattcttg aagacgaaag ggcctcgtga
     5161 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5221 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5281 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5341 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5401 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5461 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5521 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5581 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5641 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5701 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5761 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5821 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5881 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5941 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6001 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6061 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6121 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6181 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6241 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6301 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6361 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6421 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6481 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6541 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6601 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6661 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6721 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6781 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6841 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6901 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6961 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7021 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7081 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7141 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagca
     7201 agctcatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     7261 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     7321 cgtggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7381 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7441 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt
     7501 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7561 tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc
     7621 ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg
     7681 attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact
     7741 tagtagt
//
