LOCUS       sequence-360148-        7803 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 7803)
  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 7803)
  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 Cullin1
FEATURES             Location/Qualifiers
     source          1..7803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    1967..2555
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2020..2040)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2438..2449)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2630..2863
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2982..3000)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3068..3172
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3173..3241,3242..4033)
                     /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(3391..3410)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4204..4792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4693..4712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4946..4963
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5203..5224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5239..5244,5245..5262,5263..5269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5277..5293
                     /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     5282..5304
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5301..5317
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5301..5317
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5367..5386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5578..6211
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6258..6383
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6880..7113
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7298..7342
                     /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    7635..7752
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccggcgg ccgatccacc atgtcgtcaa
      601 cccggagcca gaacccccac ggcctgaagc agattggcct ggaccagatc tgggacgacc
      661 tcagagccgg catccagcag gtgtacacac ggcagagcat ggccaagtcc agatatatgg
      721 agctctacac tcatgtttat aactactgta ctagtgttca ccagtcaaac caagcacgag
      781 gagctggagt tcctccttct aagtcgaaaa aggggcagac acctggagga gctcagtttg
      841 ttggcctgga attatataaa cgacttaagg aatttttgaa gaattacttg acaaatcttc
      901 ttaaggatgg agaagatttg atggatgaga gtgtactgaa attctacact caacaatggg
      961 aagattatcg attttcaagc aaagtgctga atggaatttg tgcctacctc aatagacatt
     1021 gggttcgccg tgaatgtgac gaaggacgaa aaggaatata tgaaatctat tcgcttgcat
     1081 tggtgacttg gagagactgt ctgttcaggc cactgaataa acaggtaaca aatgctgttt
     1141 taaagctgat tgaaaaggaa aggaatggtg aaaccatcaa tacaagattg attagtggag
     1201 ttgtacagtc ttacgtggaa ttggggctga atgaagatga tgcatttgca aagggcccta
     1261 cgttaacagt gtataaagaa tcctttgaat ctcaattttt ggctgacaca gagagatttt
     1321 ataccagaga gagtactgaa ttcttgcagc agaacccagt tactgaatat atgaaaaagg
     1381 cagaggctcg tctgcttgag gaacaacgaa gagttcaggt ttaccttcat gaaagcacac
     1441 aagatgaatt agcaaggaaa tgtgaacaag tcctcattga aaaacacttg gaaattttcc
     1501 acacagaatt tcagaattta ttggatgctg acaaaaatga agatttggga cgcatgtata
     1561 atcttgtatc tagaatccag gatggcctag gagaattgaa aaaactgttg gagacacaca
     1621 ttcataatca gggtcttgca gccattgaaa agtgtggaga agctgcttta aatgacccca
     1681 aaatgtatgt acagacagtg cttgatgttc ataaaaaata caatgccctg gtaatgtctg
     1741 cattcaacaa tgacgctggc tttgtggctg ctcttgataa ggcttgtggt cgcttcataa
     1801 acaacaacgc ggttaccaag atggcccaat catccagtaa atcccctgag ttgctggctc
     1861 gatactgtga ctccttgttg aagaaaagtt ccaagaaccc agaggaggca gaactagaag
     1921 acacactcaa tcaagtgatg gttgtctaag cggcccgata gatcctaatc aacctctgga
     1981 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2041 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2101 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2161 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2221 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2281 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2341 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     2401 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2461 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2521 gacgagtcgg atctcccttt gggccgcctc cccgcctgag atcctttaag accaatgact
     2581 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     2641 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     2701 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     2761 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     2821 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     2881 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggcccggg
     2941 ttaattaagg aaagggctag atcattcttg aagacgaaag ggcctcgtga tacgcctatt
     3001 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3061 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3121 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3181 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3241 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3301 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3361 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     3421 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3481 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3541 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3601 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3661 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3721 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3781 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3841 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3901 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3961 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4021 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4081 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4141 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4201 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4261 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4321 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4381 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4441 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4501 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4561 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4621 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4681 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4741 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4801 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4861 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4921 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     4981 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagca agctcatggc
     5041 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5101 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc cgtggcacga
     5161 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5221 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5281 gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt tcacaaataa
     5341 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5401 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
     5461 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
     5521 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagttgg
     5581 aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac
     5641 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
     5701 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
     5761 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     5821 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
     5881 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
     5941 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
     6001 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
     6061 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6121 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6181 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     6241 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     6301 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     6361 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     6421 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     6481 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     6541 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     6601 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     6661 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     6721 cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     6781 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     6841 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     6901 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     6961 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     7021 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     7081 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     7141 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     7201 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     7261 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     7321 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     7381 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     7441 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     7501 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     7561 agacagatcc attcgattag tgaacggatc tcgacggtat cgccgaattc acaaatggca
     7621 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     7681 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     7741 attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggac tagtggagtt
     7801 ccg
//
