LOCUS       sequence-454055-        7436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xFLAG-tagged CUL4B in a pCMV7.1 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7436)
  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 7436)
  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-CUL4B-pCMV7.1
FEATURES             Location/Qualifiers
     source          1..7436
                     /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    3460..4082
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3659..3678)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4106..4126)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4111..4440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4291..4426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4478..4497)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4479..4497)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4511..4527)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4511..4527)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4524..4546)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4535..4551
                     /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(4559..4565,4566..4583,4584..4589))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4604..4625
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4742..4759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4913..5501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4993..5012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5672..6463,6464..6532))
                     /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     6295..6314
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6533..6637)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6664..7119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6751..6770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6961..6982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7245..7267
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7259..7276
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7260..7276
                     /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 gatgatgtca cagtcatctg gatcaggaga
      721 tgggaatgat gatgaggcta ctacctctaa agacggtggt ttttcttccc ccagtccctc
      781 agctgctgct gctgctcagg aggtcagatc tgccactgat ggtaatacca gcaccactcc
      841 gcccacctct gccaagaaga gaaagttaaa cagcagcagc agtagcagca gtaacagtag
      901 taacgagaga gaagactttg attccacctc ttcctcctct tccactcctc ctttacaacc
      961 cagggattcg gcatcccctt caacctcgtc cttctgcctg ggggtttcag tggctgcttc
     1021 cagccacgta ccgatacaga agaagctgcg ttttgaagac accctggagt ttgtagggtt
     1081 tgatgcgaag atggctgagg aatcctcctc ctcctcctcc tcatcttcac caactgctgc
     1141 aacatctcag cagcagcaac ttaaaaataa gagtatatta atctcttctg tggcttcggt
     1201 gcatcatgca aacggcctag ccaaatcttc taccaccgtc tctagctttg ctaacagcaa
     1261 acctggctct gctaagaagt tagtgatcaa gaactttaaa gataagccta aattaccaga
     1321 aaactacaca gatgaaacct ggcaaaaact gaaagaagca gtggaagcta ttcagaatag
     1381 tacttcaatt aagtacaatt tagaagaact ctaccaggct gtagaaaatc tctgttctta
     1441 caagatttct gcaaacttgt acaaacagct gagacagatc tgcgaagatc acatcaaagc
     1501 acagattcat caattcagag aggattcatt ggatagcgtt ctttttttaa agaagattga
     1561 tagatgctgg caaaaccatt gcagacaaat gatcatgatc aggagcattt ttttgtttct
     1621 ggatagaact tacgttcttc agaattcaat gctaccctcc atttgggaca tgggactgga
     1681 gttatttagg gctcatatta taagtgatca gaaagtgcag aataagacaa ttgatggcat
     1741 tcttctcttg attgagaggg aaaggaatgg tgaagcaatt gatagaagtt tacttcgaag
     1801 ccttttaagc atgctgtctg atttgcaaat ttatcaagat tcttttgaac aacgattttt
     1861 ggaagaaact aaccggctct atgcagctga aggccaaaaa ttaatgcaag aaagagaggt
     1921 tcctgaatat ctacatcatg ttaacaaacg tctagaagaa gaagcagaca gacttattac
     1981 ttacttagat cagaccaccc agaagtcatt aattgctact gtagaaaaac aacttctagg
     2041 tgaacactta acagcaattc ttcagaaagg tttaaataac ctccttgatg aaaaccgaat
     2101 tcaagatttg tctcttctgt atcagctctt cagtagagtt cgaggtggag ttcaggttct
     2161 tttgcagcag tggatcgaat atatcaaggc atttggcagc actattgtaa ttaatcctga
     2221 aaaagataaa accatggttc aagaattgct ggattttaaa gataaggttg accatataat
     2281 tgatatctgc tttctgaaga atgagaaatt tatcaatgcc atgaaagaag catttgaaac
     2341 gttcattaac aaaagaccaa ataaaccagc tgaacttata gctaagtatg tagattcaaa
     2401 acttcgtgca ggcaacaaag aagctacaga tgaagaactt gagaaaatgt tggataaaat
     2461 tatgatcata tttagattta tctatggcaa ggatgttttt gaggccttct ataagaaaga
     2521 tttagccaag cgcctgttag tcggaaagag tgcatctgta gatgctgaaa aatcaatgct
     2581 gtccaaactt aaacatgaat gcggagctgc tttcaccagc aaacttgaag gaatgtttaa
     2641 agacatggaa ctttctaaag acatcatgat tcagttcaaa cagtatatgc agaatcagaa
     2701 tgttccggga aatattgagt taactgtgaa tatcctgaca atgggctatt ggccgacata
     2761 tgtgcctatg gaagttcatt taccaccaga gatggtaaaa cttcaggaga ttttcaagac
     2821 attttaccta ggcaaacata gtggcaggaa acttcagtgg cagtcaaccc taggacactg
     2881 tgtgttaaaa gcagaattta aagagggtaa aaaggaactc caggtctctc tttttcaaac
     2941 actggtgctg ctaatgttta atgagggaga ggagttcagt ttagaagaga tcaagcaggc
     3001 aactggaata gaggatggag agttaaggag aacactgcag tcattagcct gtggcaaagc
     3061 tagagttctg gcgaaaaatc caaagggcaa agacattgaa gatggtgaca agttcatttg
     3121 taatgatgat ttcaaacata aacttttcag gataaagatc aatcaaatcc agatgaaaga
     3181 aacggttgaa gaacaagcaa gcactacaga aagagtattt caagacagac agtatcaaat
     3241 tgatgctgca attgttcgaa ttatgaagat gagaaagaca cttagccaca atctccttgt
     3301 ttcagaagtg tacaaccagt tgaaatttcc agtaaagcct gctgatctta agaagagaat
     3361 agaatcttta attgaccggg actacatgga aagagataaa gaaaatccaa accagtacaa
     3421 ctatattgca tagggtacca gtcgactcta gaggatcccg ggtggcatcc ctgtgacccc
     3481 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     3541 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     3601 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     3661 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     3721 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     3781 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     3841 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     3901 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag
     3961 caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg
     4021 agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa
     4081 ttgggtacgc ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4141 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4201 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4261 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4321 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4381 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4441 aagctcctcg aggaactgaa aaaccagaaa gttaattccc tatagtgagt cgtattaaat
     4501 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4561 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4621 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4681 attaatgaat cggccaacgc gcggagagag gcggtttgcg tattgggcgc tcttccgctt
     4741 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4801 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4861 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4921 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4981 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5041 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5101 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5161 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5221 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5281 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5341 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5401 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5461 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5521 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5581 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5641 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5701 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5761 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5821 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5881 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5941 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6001 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6061 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6121 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6181 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6241 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6301 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6361 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6421 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6481 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6541 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6601 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6661 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6721 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6781 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6841 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6901 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6961 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     7021 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     7081 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
     7141 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     7201 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     7261 taaaacgacg gccagtgcca agctgatcta tacattgaat caatattggc aattagccat
     7321 attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
     7381 tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgtt
//
