LOCUS       sequence-311396-        7001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xFLAG-tagged CUL3 in a pCMV7.1 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7001)
  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 7001)
  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-CUL3-pCMV7.1
FEATURES             Location/Qualifiers
     source          1..7001
                     /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    3025..3647
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3224..3243)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3671..3691)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3676..4005
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3856..3991
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4043..4062)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4044..4062)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4076..4092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4076..4092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4089..4111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4100..4116
                     /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(4124..4130,4131..4148,4149..4154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4169..4190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4307..4324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4478..5066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4558..4577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5237..6028,6029..6097))
                     /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     5860..5879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6098..6202)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6229..6684
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6316..6335)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6526..6547
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6810..6832
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6824..6841
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6825..6841
                     /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 gatgtcgaat ctgagcaaag gcacgggcag
      721 ccggaaggac accaagatgc ggatccgggc ctttccgatg accatggatg aaaaatatgt
      781 aaacagcatt tgggaccttc tgaaaaatgc aattcaagaa atccagcgta agaataacag
      841 tggtcttagt tttgaggagc tctatagaaa tgcatataca atggttttgc ataaacatgg
      901 agaaaagctc tacactggac taagagaagt tgttaccgaa catctcataa ataaggtgcg
      961 agaagatgta ctaaattcat tgaataacaa ctttcttcaa acgctaaatc aagcttggaa
     1021 tgatcatcaa acagctatgg tgatgattag agacatacta atgtacatgg accgtgtgta
     1081 tgtacaacaa aataatgtgg agaacgtcta caatttggga ttaattattt ttcgagatca
     1141 agttgtacgt tatgggtgta ttagggatca tctacggcaa actctattgg atatgattgc
     1201 aagagagcgg aaaggagaag tcgtagacag aggcgcaata agaaatgctt gccagatgtt
     1261 aatgatttta ggtctcgaag gaagatcagt ctatgaagaa gattttgagg ctcctttttt
     1321 ggaaatgtct gcagaatttt ttcagatgga aagccagaaa tttttagcag aaaatagtgc
     1381 ttcagtatat ataaagaaag tagaagctag aattaatgaa gaaatagaac gagtgatgca
     1441 ctgccttgac aaatcaacgg aagaaccaat tgtaaaggtg gttgaaaggg aactcatttc
     1501 caagcacatg aagactatag tagaaatgga gaattctggg ctagtacata tgttgaaaaa
     1561 tggaaagaca gaagaccttg gttgcatgta caagttattt agtcgtgtgc caaatggttt
     1621 gaaaacaatg tgtgagtgta tgagttccta tttgagggag caaggtaaag ctcttgtttc
     1681 tgaagaagga gaaggaaaga atcctgttga ctatatccag ggcttattgg atctgaagag
     1741 taggttcgat cgcttcctcc tggaatcatt caacaatgac cgtctcttta aacaaactat
     1801 tgcgggtgac tttgagtatt ttctcaacct caactccagg tctcctgaat acctctcatt
     1861 atttattgat gataagctga aaaagggagt caaagggcta acagaacaag aagtagaaac
     1921 aatattggat aaagcaatgg tcctttttag gtttatgcaa gaaaaagatg tatttgaacg
     1981 ttattataaa caacacttgg caaggagact tctcacaaat aaaagtgttt ctgatgactc
     2041 tgaaaaaaac atgatatcta agttaaagac tgaatgtgga tgtcagttca cgtcaaaact
     2101 ggaaggaatg tttagggata tgagcatctc aaacacaacg atggatgaat tcaggcaaca
     2161 tctacaggca actggtgtat ctttaggtgg tgttgatctt acagtccggg tgctcacgac
     2221 aggatattgg cccactcagt cagccacacc aaagtgcaac atcccaccag caccaagaca
     2281 tgcttttgag atattcagaa ggttctactt agccaaacac agtggtcgac agctcacact
     2341 ccagcatcat atgggttctg cagatctcaa tgccacattt tatggaccag ttaaaaagga
     2401 agatggatct gaagttggtg ttggaggtgc acaagtaact ggctctaata cacggaagca
     2461 catattgcaa gtttccactt tccagatgac catattaatg ctctttaata atagagaaaa
     2521 atacacattt gaggaaattc agcaagagac agatatccct gaaagagagc ttgttagagc
     2581 cctacagtcc ctcgcctgtg gtaaaccaac acagcgggtt cttacaaaag aacccaaatc
     2641 aaaggaaata gaaaatggtc atatatttac agttaatgat cagttcacat ccaaactaca
     2701 cagagtcaag attcaaacag ttgctgccaa acaaggtgaa tccgacccag agaggaaaga
     2761 aacaaggcag aaagtagacg acgacagaaa acatgagata gaagctgcta tagtgcggat
     2821 aatgaaatct agaaagaaga tgcagcacaa tgttctagta gcggaggtaa ctcagcagtt
     2881 gaaggcgcga ttcttaccaa gtccagttgt tattaagaaa cgtattgaag gacttattga
     2941 gagagaatat ttggcacgaa cacctgagga tcgcaaagta tacacatatg tagcataagg
     3001 taccagtcga ctctagagga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     3061 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3121 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3181 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3241 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3301 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3361 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3421 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3481 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     3541 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     3601 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     3661 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3721 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3841 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3901 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3961 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa
     4021 ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat taaattcgta atcatgtcat
     4081 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4141 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4201 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4261 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4321 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4381 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4441 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4501 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4561 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4621 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4681 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4741 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4801 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4861 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4921 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4981 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5041 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5101 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5161 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5221 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5281 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5341 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5401 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5461 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5521 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5581 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5641 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5701 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5761 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5821 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5881 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5941 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6001 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6061 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6121 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6181 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     6241 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6301 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6361 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6421 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6481 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6541 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6601 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6661 acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg
     6721 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6781 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     6841 tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta
     6901 tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat
     6961 gtacatttat attggctcat gtccaatatg accgccatgt t
//
