LOCUS       sequence-311400-        6997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xFLAG-tagged CUL4A in a pCMV7.1 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6997)
  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 6997)
  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-CUL4A-pCMV7.1
FEATURES             Location/Qualifiers
     source          1..6997
                     /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    3021..3643
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3220..3239)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3667..3687)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3672..4001
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3852..3987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4039..4058)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4040..4058)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4072..4088)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4072..4088)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4085..4107)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4096..4112
                     /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(4120..4126,4127..4144,4145..4150))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4165..4186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4303..4320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4474..5062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4554..4573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5233..6024,6025..6093))
                     /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     5856..5875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6094..6198)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6225..6680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6312..6331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6522..6543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6806..6828
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6820..6837
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6821..6837
                     /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 gatatggcgg acgaggcccc
      721 gcggaagggc agcttctcgg cgctcgtggg ccgcaccaac ggcctcacca agcccgcggc
      781 cctggccgcc gcgcccgcca agccgggggg cgcgggcggc tccaagaagc tggtcatcaa
      841 gaacttccga gacagacctc ggctgcccga caactacacg caggacacgt ggcggaagct
      901 gcacgaggcg gtgcgggccg tgcagagcag cacctccatc aggtacaacc tcgaggagct
      961 ctaccaggct gtggaaaatc tctgttctca caaagtctcc ccaatgctct acaagcaact
     1021 gcgtcaggcc tgtgaagacc acgtccaggc acagatcctt ccgtttagag aagactcact
     1081 agatagtgtt ttatttttaa agaagattaa cacgtgctgg caggaccact gcagacaaat
     1141 gatcatgatc agaagcatct tcctgttctt ggaccgcacc tatgtgctgc agaactccac
     1201 gctgccctcc atctgggata tgggattaga actgtttaga acccatatta ttagtgataa
     1261 aatggttcag agtaaaacca ttgatggaat cctactgctg atcgagcgcg agaggagcgg
     1321 cgaggccgtg gaccggagcc tgttgcggag cctcctgggc atgctgtctg acctgcaggt
     1381 gtataaagat tcatttgaac tgaaattttt ggaagagact aattgcttat atgctgccga
     1441 aggccaaagg ttaatgcagg aaagagaggt tccagaatat cttaaccatg taagtaaacg
     1501 cttagaggaa gagggagaca gagtaatcac ttacttggac cacagcacac agaaaccact
     1561 gattgcttgt gtggagaaac agctattagg agaacattta acagcaattc tgcagaaagg
     1621 gctcgaccac ttactggatg agaacagagt gccggacctc gcacagatgt accagctgtt
     1681 cagccgggtg aggggcgggc agcaggcgct gctgcagcac tggagcgagt acatcaagac
     1741 ttttggaaca gcgatcgtaa tcaatcctga gaaagacaaa gacatggtcc aagacctgtt
     1801 ggacttcaag gacaaggtgg accacgtgat cgaggtctgc ttccagaaga atgagcggtt
     1861 cgtcaacctg atgaaggagt cctttgagac gttcatcaac aagagaccca acaagcctgc
     1921 agaactgatc gcaaagcatg tggattcaaa gttaagagca ggcaacaaag aagccacaga
     1981 cgaggagctg gagcggacgt tggacaagat catgatcctg ttcaggttta tccacggtaa
     2041 agatgtcttt gaagcatttt ataaaaaaga tttggcaaaa agactccttg ttgggaaaag
     2101 tgcctcagtc gatgctgaaa agtctatgtt gtcaaagctc aagcatgagt gcggtgcagc
     2161 cttcaccagc aagctggaag gcatgttcaa ggacatggag ctttcgaagg acatcatggt
     2221 tcatttcaag cagcatatgc agaatcagag tgactcaggc cctatagacc tcacagtgaa
     2281 catactcaca atgggctact ggccaacata cacgcccatg gaagtgcact taaccccaga
     2341 aatgattaaa cttcaggaag tatttaaggc attttatctt ggaaagcaca gtggtcgaaa
     2401 acttcagtgg caaactactt tgggacatgc tgttttaaaa gcggagttta aagaagggaa
     2461 gaaggaattc caggtgtccc tcttccagac actggtgctc ctcatgttca acgagggaga
     2521 tggcttcagc tttgaggaga taaaaatggc cacggggata gaggatagtg aattgcgcag
     2581 aacgctgcag tccctggcct gtggcaaagc acgtgtgctg attaaaagtc ccaaaggaaa
     2641 ggaagtggaa gatggagaca agttcatttt taatggagag ttcaagcaca agttgtttag
     2701 aataaagatc aatcaaattc agatgaagga aactgttgag gaacaggtta gcaccactga
     2761 gagagtgttt caggatagac aatatcagat tgatgctgct atcgtcagaa taatgaagat
     2821 gagaaagact cttggtcata atcttctagt ttctgaatta tataatcagc tgaaatttcc
     2881 agtaaagcct ggagatttga aaaagagaat tgaatctctg atagacagag actatatgga
     2941 gagagacaaa gacaatccga atcagtacca ctacgtggcc tgaatctgca gaattccacc
     3001 acactggact agtggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     3061 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     3121 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     3181 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     3241 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     3301 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     3361 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     3421 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     3481 ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac
     3541 agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg
     3601 tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt
     3661 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3721 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3781 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3841 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3901 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3961 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga
     4021 aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct
     4081 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4141 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4201 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4261 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4321 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4381 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4981 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5041 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5101 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5161 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5221 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5281 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5341 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5401 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5461 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5521 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5581 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5641 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5701 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5761 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5821 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5881 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5941 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6001 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6061 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6121 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6181 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     6241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6301 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6361 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6421 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6541 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6601 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6661 aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6721 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6781 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     6841 aagctgatct atacattgaa tcaatattgg caattagcca tattagtcat tggttatata
     6901 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     6961 atttatattg gctcatgtcc aatatgaccg ccatgtt
//
