LOCUS       sequence-311405-        7060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xFLAG-tagged CUL5 in a pCMV7.1 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7060)
  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 7060)
  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-CUL5-pCMV7.1
FEATURES             Location/Qualifiers
     source          1..7060
                     /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    3084..3706
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3283..3302)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3730..3750)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3735..4064
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3915..4050
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4102..4121)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4103..4121)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4135..4151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4135..4151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4148..4170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4159..4175
                     /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(4183..4189,4190..4207,4208..4213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4228..4249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4366..4383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4537..5125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4617..4636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5296..6087,6088..6156))
                     /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     5919..5938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6157..6261)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6288..6743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6375..6394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6585..6606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6869..6891
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6883..6900
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6884..6900
                     /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 gaattcatcg atagatctga tatcggtacc
      721 agtcgacatg gcgacgtcta atctgttaaa gaataaaggt tctcttcagt ttgaagacaa
      781 atgggatttt atgcgcccga ttgttttgaa gcttttacgc caggaatctg ttacaaaaca
      841 gcagtggttt gatctgtttt cggatgtgca tgcagtctgt ctttgggatg ataaaggccc
      901 agcaaaaatt catcaggctt taaaagaaga tattcttgag tttattaagc aggcacaggc
      961 acgagtactg agccatcaag atgatacggc tttgctaaaa gcatatattg ttgaatggcg
     1021 aaagttcttt acacaatgtg atattttacc aaaacctttt tgtcaactag agattacttt
     1081 aatgggtaaa cagggcagca ataaaaaatc aaatgtggaa gacagtattg ttcgaaagct
     1141 tatgcttgat acatggaatg agtcaatctt ttcaaacata aaaaacagac tccaagatag
     1201 tgcaatgaag ctggtacatg ctgagagatt gggagaagct tttgattctc agctggttat
     1261 tggagtaaga gaatcctatg ttaacctttg ttctaatcct gaggataaac ttcaaattta
     1321 tagggacaat tttgagaagg catacttgga ttcaacagag agattttata gaacacaagc
     1381 accctcgtat ttacaacaaa atggtgtaca gaattatatg aaatatgcag atgctaaatt
     1441 aaaagaagaa gaaaaacgag cactacgtta tttagaaaca agacgagaat gtaactccgt
     1501 tgaagcactc atggaatgct gtgtaaatgc cctggtgaca tcatttaaag agactatctt
     1561 agctgagtgc caaggcatga tcaagagaaa tgaaactgaa aaattacatt taatgttttc
     1621 attgatggac aaagttccta atggtataga gccaatgttg aaagacttgg aggaacatat
     1681 cattagtgct ggcctggcag atatggtagc agctgctgaa actattacta ctgactctga
     1741 gaaatacgtt gagcagttac ttacactatt taatagattt agtaaactcg tcaaagaagc
     1801 ttttcaagat gatccacgat ttcttactgc aagagataag gcgtataaag cagttgttaa
     1861 tgatgctacc atatttaaac ttgaattacc tttgaagcag aagggggtgg gattaaaaac
     1921 tcagcctgaa tcaaaatgcc ctgagctgct tgccaattac tgtgacatgt tgctaagaaa
     1981 aacaccatta agcaaaaaac taacctctga agagattgaa gcaaagctta aagaagtgct
     2041 cttggtactt aagtatgtac agaacaaaga tgtttttatg aggtatcata aagctcattt
     2101 gacacgacgt cttatattag acatctctgc cgatagtgaa attgaagaaa acatggtaga
     2161 gtggctaaga gaagttggta tgccagcgga ttatgtaaac aagcttgcta gaatgtttca
     2221 ggacataaaa gtatctgaag atttgaacca agcttttaag gaaatgcaca aaaataataa
     2281 attggcatta ccagctgatt cagttaatat aaaaattctg aatgctggcg cctggtcaag
     2341 aagttctgag aaagtctttg tctcacttcc tactgaactg gaggacttga taccggaagt
     2401 agaagaattc tacaaaaaaa atcatagtgg tagaaaatta cattggcatc atctcatgtc
     2461 aaatggaatt ataacattta agaatgaagt tggtcaatat gatttggagg taaccacgtt
     2521 tcagctcgct gtattgtttg catggaacca aagacccaga gagaaaatca gctttgaaaa
     2581 tcttaagctt gcaactgaac tccctgatgc tgaacttagg aggactttat ggtctttagt
     2641 agctttccca aaactcaaac ggcaagtttt gttgtatgaa cctcaagtca actcacccaa
     2701 agactttaca gaaggtaccc tcttctcagt gaaccaggag ttcagtttaa taaaaaatgc
     2761 aaaggttcag aaaaggggta aaatcaactt gattggacgt ttgcagctca ctacagaaag
     2821 gatgagagaa gaagagaatg aaggaatagt tcaactacga atactaagaa cccaggaagc
     2881 tatcatacaa ataatgaaaa tgagaaagaa aattagtaat gctcagctgc agactgaatt
     2941 agtagaaatt ttgaaaaaca tgttcttgcc acaaaagaaa atgataaaag agcaaataga
     3001 gtggctaata gagcacaaat acatcagaag agatgaatct gatatcaaca ctttcatata
     3061 tatggcataa tctagaggat cccgggtggc atccctgtga cccctcccca gtgcctctcc
     3121 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     3181 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     3241 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     3301 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     3361 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     3421 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     3481 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     3541 ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga
     3601 cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca
     3661 ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag
     3721 cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3781 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3841 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3901 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3961 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4021 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ctcgaggaac
     4081 tgaaaaacca gaaagttaat tccctatagt gagtcgtatt aaattcgtaa tcatgtcata
     4141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     6301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     6361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6421 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     6481 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     6541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     6601 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     6661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     6721 caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg
     6781 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     6841 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     6901 gccaagctga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
     6961 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     7021 tacatttata ttggctcatg tccaatatga ccgccatgtt
//
