LOCUS       sequence-311409-        9791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xFLAG-tagged CUL7 in a pCMV7.1 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9791)
  AUTHORS   Sweeney MA, Iakova P, Maneix L, Shih FY, Cho HE, Sahin E, Catic A
  TITLE     The ubiquitin ligase Cullin-1 associates with chromatin and 
            regulates transcription of specific c-MYC target genes.
  JOURNAL   Sci Rep. 2020 Aug 18;10(1):13942. doi: 10.1038/s41598-020-70610-0.
  PUBMED    32811853
REFERENCE   2  (bases 1 to 9791)
  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"; journalName: "Sci Rep."; 
            date: "2020-08-18"; volume: "10(1):13942. doi"; pages: " 
            10.1038/s41598-020-70610-0"
COMMENT     Sequence Label: 3xFLAG-CUL7-pCMV7.1
FEATURES             Location/Qualifiers
     source          1..9791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    5815..6437
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6014..6033)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6461..6481)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6466..6795
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6646..6781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6833..6852)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6834..6852)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6866..6882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6866..6882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6879..6901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6890..6906
                     /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)."
     promoter        complement(join(6914..6920,6921..6938,6939..6944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6959..6980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7097..7114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7268..7856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7348..7367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8027..8818,8819..8887))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8650..8669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8888..8992)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9019..9474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9106..9125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9316..9337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9600..9622
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9614..9631
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9615..9631
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaagc ttgcggccgc gatggtggga gaactccgct acagggaatt
      721 cagggtgccc ctggggcccg gcttacatgc ctatcctgat gagctgatcc gccagcgcgt
      781 gggccatgat gggcatcctg agtaccagat ccgttggctc atcctgcggc gtggcgatga
      841 gggggacggg ggctctggcc aagtggactg caaggctgag cacatcctgc tgtggatgtc
      901 caaggatgag atctatgcca actgccacaa gatgctgggc gaggatggcc aggtcatcgg
      961 gccctcccag gagtctgcag gggaggttgg ggccctggac aaatctgtgc tggaggagat
     1021 ggaaaccgac gtgaagtccc tcattcagag agcccttcgg cagctggagg agtgtgtggg
     1081 cactatccct cctgctcctc tacttcacac tgtccacgtg ctcagcgcct atgccagcat
     1141 tgagcccctc actggagtat tcaaggaccc aagggtcctg gacttgctca tgcacatgtt
     1201 gagtagtcct gattatcaga ttcgctggag tgcaggccgg atgatacaag ccctgtcctc
     1261 ccatgacgct gggacccgga ctcagatcct tctgtcactg agccaacaag aagccattga
     1321 gaaacacctg gattttgaca gccgctgtgc tctgctagca ctgtttgcac aggccacgct
     1381 ctctgaacac cccatgtctt tcgagggcat tcagctacca caggtcccag gaagggtgct
     1441 cttctccctg gtgaagcggt atttgcatgt cacctcgctc ctggatcagc tgaacgacag
     1501 tgctgcggag ccaggagccc agaacacctc tgctcctgag gagttgagtg gggagagggg
     1561 tcaactggag ctggagttca gtatggccat gggcaccctg atctcggagc tggtgcaagc
     1621 catgcgctgg gaccaggcct cagacagacc aaggagctca gcacggtccc ccggttccat
     1681 cttccagcct cagctggcag atgtgagccc agggctcccc gctgcccagg ctcagccctc
     1741 cttcaggagg tcaagacgtt ttcgccctcg ttctgagttc gcaagtggca atacctatgc
     1801 tttgtatgtg cgggacacac tgcagccggg gatgcgagtg cggatgctgg atgattatga
     1861 ggagatcagt gccggggatg agggcgagtt tcggcagagc aacaacggtg tgcctcctgt
     1921 gcaggtattt tgggagtcaa caggccgcac ctattgggtg cactggcaca tgctggagat
     1981 cttgggcttt gaggaagaca ttgaggacat ggttgaggct gatgagtacc aaggggcagt
     2041 ggccagtaga gtcctgggta gagccctgcc tgcctggcgc tggaggccca tgacagaact
     2101 ctatgctgtg ccttatgtgc tgcctgagga tgaggacact gaggagtgtg aacacctgac
     2161 cctggctgag tggtgggaac tcctcttctt catcaagaag ctggatggac ctgaccatca
     2221 ggaggttctc cagatcctcc aggagaacct agatggggag attctggatg atgagatcct
     2281 agctgaactg gccgtgccca tagaattggc ccaggacttg ctgctgactc tgccacagcg
     2341 actcaatgac agtgccctca gggacctgat caactgccat gtctacaaga agtatgggcc
     2401 tgaagcccta gcagggaacc aagcctaccc atcccttcta gaagcccaag aagatgtcct
     2461 cctgctagac gcgcaggccc aggctaagga ctcagaagat gcagccaaag tggaagcaaa
     2521 agaaccccca tctcagagtc ccaacactcc cctgcagcgt ctggtggagg gttatggtcc
     2581 agctgggaaa atcctcctgg atctagagca agccctcagc tcagagggga cccaggagaa
     2641 caaggtcaag ccactcctgc tgcagctgca gcggcagccg cagcccttcc tggcactgat
     2701 gcagagcctg gacactccgg agactaacag gaccctgcac ctgactgtgc tgagaatcct
     2761 gaagcagctg gtggacttcc ccgaggcact gctgctcccc tggcacgagg ccgtggatgc
     2821 ctgcatggcc tgcctgcggt ccccaaacac tgatcgagag gtgctccagg aactgatttt
     2881 cttcctgcac cgcctgacct cagtgagcag ggactatgcc gtggtgctga atcagctggg
     2941 agcaagagac gctatctcca aggccctgga aaagcacctg ggaaagctgg agctggctca
     3001 ggagctgcgg gacatggtgt tcaagtgtga gaagcatgcc cacctctacc gcaaactcat
     3061 caccaacatc ctgggaggct gcatccagat ggtgctgggc cagatcgaag accacagacg
     3121 aacccaccgg cccatcaaca tccctttctt tgatgtgttc ctcagatacc tgtgccaggg
     3181 ctccagtgtg gaagtgaagg aggacaagtg ctgggagaag gtggaggtgt cctccaaccc
     3241 gcaccgggcc agcaagctga cggaccacaa ccccaagacc tattgggagt ccaacggcag
     3301 cgccggctcc cactacatca ccctgcacat gcgccggggc atcctcatca ggcaactgac
     3361 tctgcttgtg gctagtgagg actcgagtta catgccggcc cgagtggtgg tgtgcggggg
     3421 tgatagcact agctctcttc acacggaact caactcggtg aatgtgatgc cctctgccag
     3481 ccgggtgatc ctcctggaga acctgacccg cttctggccc atcatccaga tccgcataaa
     3541 gcgctgccag cagggtggca ttgatacgcg cattcggggg ttagagatcc taggccccaa
     3601 gcccacgttc tggccagtgt tccgggagca gctctgtcgt cacacacgcc tcttctacat
     3661 ggttcgggca caggcctgga gccaggacat ggcagaggac cgcaggagcc tcctgcacct
     3721 gagttctaga ctcaacggtg ctctgcgcca ggagcagaat tttgctgacc gcttcctccc
     3781 tgatgacgag gctgcccaag ctctgggcaa gacctgctgg gaggccctgg tcagccccgt
     3841 ggtgcagaac atcacctccc ctgatgagga tggcattagc cccctgggtt ggctgctgga
     3901 ccagtacctg gagtgtcagg aagctgtctt caacccccag agccgcggcc cagctttctt
     3961 ctcgcgggtg cgccgtctca ctcacctgct ggtgcatgtc gagccctgtg aggcaccccc
     4021 tcctgtggtg gccactcctc ggcccaaagg cagaaacaga agccacgact ggagctcctt
     4081 ggctacccgg ggccttccaa gcagcatcat gagaaacctg acgcgctgtt ggcgggccgt
     4141 ggtggagaag caggtgaaca attttctgac ctcatcctgg cgggatgatg actttgtgcc
     4201 acgctactgt gagcacttta atattctgca gaactcaagc tctgaactgt ttgggcctcg
     4261 ggcagccttc ttgctggcgc tgcaaaatgg ctgtgcggga gccttgctga agctcccttt
     4321 tctcaaagct gcccacgtga gtgagcagtt cgcccggcac attgaccagc agatccaggg
     4381 cagccggatc ggtggagccc aggaaatgga gaggctggca cagctgcagc aatgcctgca
     4441 agctgtcctg attttctccg gcttggagat agccaccact tttgagcatt attaccagca
     4501 ctacatggcg gaccgtctcc tgggcgtggt ctcgagctgg ctggaggggg ccgtgctgga
     4561 gcagatcggt ccctgcttcc ccaaccgcct cccccagcag atgttgcaga gcctgagcac
     4621 ctctaaggag ctgcagcgcc agttccacgt ctaccagctc cagcagctgg atcaggaact
     4681 cctgaagctg gaggatacag agaagaaaat acaggtgggc cttggggcca gtggcaagga
     4741 gcacaagagc gagaaggaag aggaagctgg ggcagcagca gtggtggatg tggcggaggg
     4801 agaggaggaa gaggaggaga atgaggacct ctactatgaa ggggcaatgc cagaagtgtc
     4861 tgtgcttgtc ctgtcccgac actcctggcc tgttgcctca atctgccaca cactgaaccc
     4921 cagaacctgc ctgccctcct acctgagggg cactttgaac agatactcca acttctacaa
     4981 caagagtcag agccaccctg cccttgagcg aggctcacag aggcgactgc agtggacgtg
     5041 gctgggctgg gctgagctgc agtttgggaa ccagaccctg catgtgtcca ccgtgcagat
     5101 gtggctactg ctgtatctca acgacctgaa ggcggtctct gtggagagtc tgctggcgtt
     5161 ctcagggctc tccgcagaca tgctcaatca ggcgattggg cccctcacct cttcaagagg
     5221 ccccctggac cttcacgagc aaaaggatat accaggaggg gtcctcaaga ttcgagatgg
     5281 cagcaaggaa cccaggtcga gatgggacat tgtgcggctc atcccacctc agacgtacct
     5341 gcaagctgag ggtgaagacg gccagaactt ggagaagaga cggaatcttc tgaactgcct
     5401 catcgtccga atcctcaagg cccatggaga tgaggggctg cacattgacc agcttgtctg
     5461 tctggtgctg gaggcttggc agaagggccc gtgtcctccc aggggtttgg tcagcagcct
     5521 tggtaagggg tctgcatgca gcagcactga cgtcctctcc tgcatcctac acctcctggg
     5581 caagggcacg ctgagacgcc atgacgaccg gccccaggtg ctgtcctatg cagtccctgt
     5641 gactgtcatg gagcctcaca ctgagtccct gaacccaggc tcctcaggcc ccaacccacc
     5701 cctcaccttc cataccctac agattcgctc ccggggtgtg ccctatgcct cctgcactgc
     5761 cacccagagc ttctctacct tccggtaggg taccagtcga ctctagagga tcccgggtgg
     5821 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     5881 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     5941 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     6001 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     6061 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     6121 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     6181 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     6241 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta
     6301 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa
     6361 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag
     6421 tagatgcctg ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg
     6481 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6541 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6601 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6661 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6721 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6781 ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag
     6841 tgagtcgtat taaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
     6901 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6961 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7021 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7081 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7141 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7201 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7261 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7321 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7381 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7441 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7501 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7561 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7621 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7681 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7741 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7801 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8161 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8341 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8641 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8881 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9001 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     9061 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     9121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     9181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     9241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     9301 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     9361 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     9421 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc
     9481 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9541 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9601 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg atctatacat tgaatcaata
     9661 ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg gctattggcc
     9721 attgcatacg ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg
     9781 accgccatgt t
//
