LOCUS       sequence-311391-        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xFLAG-tagged CUL2 in a pCMV7.1 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  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 6955)
  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-CUL2-pCMV7.1
FEATURES             Location/Qualifiers
     source          1..6955
                     /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    2979..3601
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3178..3197)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3625..3645)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3630..3959
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3810..3945
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3997..4016)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3998..4016)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4030..4046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4030..4046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4043..4065)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4054..4070
                     /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(4078..4084,4085..4102,4103..4108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4123..4144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4261..4278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4432..5020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4512..4531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5191..5982,5983..6051))
                     /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     5814..5833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6052..6156)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6183..6638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6270..6289)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6480..6501
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6764..6786
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6778..6795
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6779..6795
                     /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 cactgtgctg gatatgtctt tgaaaccaag
      721 agtagtagat tttgatgaaa catggaacaa acttttgacg acaataaaag ccgtggtcat
      781 gttggaatac gtcgaaagag caacatggaa tgaccgtttc tcagatatct atgctttatg
      841 tgtggcctat cctgaacccc ttggagaaag actttataca gaaactaaga tttttttgga
      901 aaatcatgtt cggcatttgc ataagagagt tttggagtca gaagaacaag tacttgttat
      961 gtatcatagg tactgggaag aatacagcaa gggtgcagac tatatggact gcttatatag
     1021 gtatctcaac acccagttta ttaaaaagaa taaattaaca gaagcggacc ttcagtatgg
     1081 ctatggtggt gtagatatga atgaaccact tatggaaata ggagagctag cattggatat
     1141 gtggaggaaa ttgatggttg aaccacttca ggccatcctt atccgaatgc tgctccgaga
     1201 aatcaaaaat gatcgtggtg gagaagaccc aaaccagaaa gtaatccatg gggttattaa
     1261 ctcctttgtt catgttgaac agtataagaa aaaattcccc ttaaagtttt atcaggaaat
     1321 ttttgagtct ccctttctga ctgaaacagg agagtattac aaacaagaag cttcaaattt
     1381 attacaagaa tcaaactgct cacagtatat ggaaaaggtt ctaggtagat taaaagatga
     1441 agaaattcga tgtcgaaaat acctacatcc aagttcatat actaaggtga ttcatgaatg
     1501 tcaacaacga atggtagcag accacttaca gtttttacat gcagaatgtc ataatataat
     1561 tcgacaagag aaaaaaaatg acatggcaaa tatgtacgtc ttactccgtg ctgtgtccac
     1621 tggtttacct catatgattc aggagctgca aaaccacatc catgatgagg gccttcgagc
     1681 aaccagcaac cttactcagg aaaacatgcc aacactattt gtggagtcag ttttggaagt
     1741 gcatggtaaa tttgttcagc ttatcaacac tgttttgaat ggtgatcagc attttatgag
     1801 tgcgttggat aaggccctta cgtcagttgt aaattacaga gaacctaagt ctgtttgcaa
     1861 agcacctgaa ctgcttgcta agtactgtga caacttactg aagaagtcag cgaaagggat
     1921 gacagagaat gaagtggaag acaggctcac gagcttcatc acagtgttca aatacattga
     1981 tgacaaggac gtctttcaaa agttctacgc aagaatgctg gcaaaacgtt taattcatgg
     2041 gttatccatg tctatggact ctgaagaagc catgatcaac aaattaaagc aagcctgtgg
     2101 ttatgagttt accagcaagc tacatcggat gtatacagat atgagtgtca gcgctgatct
     2161 caacaataag ttcaacaatt ttatcaaaaa ccaagacaca gtaatagatt tgggaattag
     2221 ttttcaaata tatgttctac aggctggtgc gtggcctctt actcaggctc cttcatctac
     2281 gtttgcaatt ccccaggaat tagaaaaaag tgtacagatg tttgaattat tttatagcca
     2341 acatttcagt ggaaggaaac ttacatggtt acattatctg tgtacaggtg aagttaaaat
     2401 gaactatttg ggcaaaccat atgtagccat ggttacaaca taccaaatgg cagttcttct
     2461 tgcctttaac aacagtgaaa ctgtcagtta taaagagctt caggacagca ctcagatgaa
     2521 tgaaaaggaa ctgacaaaaa caatcaaatc attacttgat gtgaaaatga ttaaccatga
     2581 ttcagaaaag gaagatattg atgcagaatc ttcgttttca ttaaatatga actttagcag
     2641 taaaagaaca aaatttaaaa ttactacatc aatgcagaaa gacacaccac aagaaatgga
     2701 gcagactaga agtgcagttg atgaggaccg gaaaatgtat ctccaagctg ctatagttcg
     2761 tatcatgaaa gcacgaaaag tgcttcggca caatgccctt attcaagagg tgattagcca
     2821 gtcaagagct aggtttaatc ccagtatcag catgattaag aagtgtattg aagttctgat
     2881 agacaaacaa tacatagaac gcagccaggc gtcggcagat gaatacagct acgtcgcgtg
     2941 aatctgcaga attccaccac actggactag tggatcccgg gtggcatccc tgtgacccct
     3001 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     3061 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     3121 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     3181 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     3241 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     3301 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     3361 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     3421 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc
     3481 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga
     3541 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat
     3601 tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3661 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3781 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3841 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3901 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3961 agctcctcga ggaactgaaa aaccagaaag ttaattccct atagtgagtc gtattaaatt
     4021 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4081 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4141 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4201 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4261 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4321 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4381 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4441 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4501 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4561 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4621 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4681 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4741 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4801 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4861 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4921 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4981 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5041 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5101 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5221 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5281 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5341 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5401 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5461 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5521 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5581 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5641 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5701 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5761 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5821 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5881 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5941 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6001 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6061 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6121 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6181 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6241 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6301 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6361 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6421 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6481 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6541 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6601 taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc
     6661 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     6721 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     6781 aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca attagccata
     6841 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     6901 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
