LOCUS       sequence-311389-        7025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xFLAG-tagged CUL1 in a pCMV7.1 vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7025)
  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 7025)
  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-CUL1-pCMV7.1
FEATURES             Location/Qualifiers
     source          1..7025
                     /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    3049..3671
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3248..3267)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3695..3715)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3700..4029
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3880..4015
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4067..4086)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4068..4086)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4100..4116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4100..4116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4113..4135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4124..4140
                     /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(4148..4154,4155..4172,4173..4178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4193..4214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4331..4348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4502..5090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4582..4601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5261..6052,6053..6121))
                     /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     5884..5903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6122..6226)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6253..6708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6340..6359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6550..6571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6834..6856
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6848..6865
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6849..6865
                     /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 gatgtcgtca acccggagcc agaaccccca
      721 cggcctgaag cagattggcc tggaccagat ctgggacgac ctcagagccg gcatccagca
      781 ggtgtacaca cggcagagca tggccaagtc cagatatatg gagctctaca ctcatgttta
      841 taactactgt actagtgttc accagtcaaa ccaagcacga ggagctggag ttcctccttc
      901 taagtcgaaa aaggggcaga cacctggagg agctcagttt gttggcctgg aattatataa
      961 acgacttaag gaatttttga agaattactt gacaaatctt cttaaggatg gagaagattt
     1021 gatggatgag agtgtactga aattctacac tcaacaatgg gaagattatc gattttcaag
     1081 caaagtgctg aatggaattt gtgcctacct caatagacat tgggttcgcc gtgaatgtga
     1141 cgaaggacga aaaggaatat atgaaatcta ttcgcttgca ttggtgactt ggagagactg
     1201 tctgttcagg ccactgaata aacaggtaac aaatgctgtt ttaaagctga ttgaaaagga
     1261 aaggaatggt gaaaccatca atacaagatt gattagtgga gttgtacagt cttacgtgga
     1321 attggggctg aatgaagatg atgcatttgc aaagggccct acgttaacag tgtataaaga
     1381 atcctttgaa tctcaatttt tggctgacac agagagattt tataccagag agagtactga
     1441 attcttgcag cagaacccag ttactgaata tatgaaaaag gcagaggctc gtctgcttga
     1501 ggaacaacga agagttcagg tttaccttca tgaaagcaca caagatgaat tagcaaggaa
     1561 atgtgaacaa gtcctcattg aaaaacactt ggaaattttc cacacagaat ttcagaattt
     1621 attggatgct gacaaaaatg aagatttggg acgcatgtat aatcttgtat ctagaatcca
     1681 ggatggccta ggagaattga aaaaactgtt ggagacacac attcataatc agggtcttgc
     1741 agccattgaa aagtgtggag aagctgcttt aaatgacccc aaaatgtatg tacagacagt
     1801 gcttgatgtt cataaaaaat acaatgccct ggtaatgtct gcattcaaca atgacgctgg
     1861 ctttgtggct gctcttgata aggcttgtgg tcgcttcata aacaacaacg cggttaccaa
     1921 gatggcccaa tcatccagta aatcccctga gttgctggct cgatactgtg actccttgtt
     1981 gaagaaaagt tccaagaacc cagaggaggc agaactagaa gacacactca atcaagtgat
     2041 ggttgtcttc aagtacatag aagacaaaga cgtatttcag aagttctatg cgaagatgct
     2101 cgccaagagg ctcgtccacc agaacagtgc aagtgacgat gccgaagcca gcatgatctc
     2161 caagttaaag caagcttgcg ggttcgagta cacctctaaa cttcagcgca tgtttcaaga
     2221 cattggcgtg agcaaagatc tgaacgagca attcaaaaag cacttgacaa actcagaacc
     2281 cctagacttg gatttcagca ttcaagtgct gagctccggg tcctggccct tccagcagtc
     2341 ttgtacattt gccttgccgt cagagttgga acgtagttat cagcgattca cagctttcta
     2401 cgccagccgc cacagtggcc gaaaattgac gtggttatat cagttgtcta aaggagaatt
     2461 ggtaactaac tgcttcaaaa acagatatac tttgcaggcg tcgacattcc agatggctat
     2521 cctgcttcag tacaacacgg aagatgccta cactgtgcag cagctgaccg acagcactca
     2581 aattaaaatg gacattttgg cgcaagtttt acagatttta ttaaagtcga agctattggt
     2641 cttggaagat gaaaatgcaa atgttgatga ggtggaattg aagccagata ccttaataaa
     2701 attatatctt ggttataaaa ataagaaatt aagggttaac atcaatgtgc caatgaaaac
     2761 cgaacagaag caggaacaag aaaccacaca caaaaacatc gaggaagacc gcaaactact
     2821 gattcaggcg gccatcgtga gaatcatgaa gatgaggaag gttctgaaac accagcagtt
     2881 acttggcgag gtcctcactc agctgtcctc caggttcaaa cctcgagtcc ctgtgatcaa
     2941 gaaatgcatt gacattctaa ttgagaaaga atatttggag cgagtggatg gtgaaaagga
     3001 cacctacagt tacttggctt aaggtaccag tcgactctag aggatcccgg gtggcatccc
     3061 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     3121 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     3181 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     3241 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     3301 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     3361 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     3421 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     3481 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
     3541 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
     3601 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
     3661 cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga
     3721 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3781 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3841 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3901 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3961 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4021 ttttgcaaaa agctcctcga ggaactgaaa aaccagaaag ttaattccct atagtgagtc
     4081 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4141 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4201 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4261 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4321 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4381 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4441 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4501 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4621 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4681 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4741 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4801 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4861 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4921 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4981 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5041 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5101 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5161 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5221 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6241 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6301 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6421 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6541 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6601 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6661 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     6721 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6781 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6841 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     6901 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     6961 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     7021 atgtt
//
