LOCUS       sequence-254717-        8175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8175)
  AUTHORS   Ohta T, Michel JJ, Schottelius AJ, Xiong Y
  TITLE     ROC1, a homolog of APC11, represents a family of cullin partners 
            with an associated ubiquitin ligase activity.
  JOURNAL   Mol Cell. 1999 Apr . 3(4):535-41.
  PUBMED    10230407
REFERENCE   2  (bases 1 to 8175)
  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: "Mol Cell. 
            1999 Apr . 3(4):535-41."
COMMENT     Sequence Label: pcDNA3-myc-CUL3
FEATURES             Location/Qualifiers
     source          1..8175
                     /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"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             663..692
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             699..728
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             735..764
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3494..3512)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3495..3512)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3532..3549)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3538..3762
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3808..4236
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3895..3914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4105..4126
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4245..4265)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4250..4579
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4430..4565
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4492..4511
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4646..5440
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4700..4719)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5310..5329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5614..5735
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5651..5670)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5705..5724
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5784..5800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5784..5800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5797..5819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5808..5824
                     /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(5832..5838,5839..5856,5857..5862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5877..5898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6015..6032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6186..6774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6266..6285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6945..7736,7737..7805))
                     /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     7568..7587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7806..7910)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7985..8004)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagcta
      661 tggagcaaaa gctcatctca gaggaagatc tggaattaga gcaaaagctc atctcagagg
      721 aagatctgga attagagcaa aagctcatct cagaggaaga tctggaattc aggatcctga
      781 tatccgcggt accgagcacc atgtcgaatc tgagcaaagg cacgggcagc cggaaggaca
      841 ccaagatgcg gatccgggcc tttccgatga ccatggatga aaaatatgta aacagcattt
      901 gggaccttct gaaaaatgca attcaagaaa tccagcgtaa gaataacagt ggtcttagtt
      961 ttgaggagct ctatagaaat gcatatacaa tggttttgca taaacatgga gaaaagctct
     1021 acactggact aagagaagtt gttaccgaac atctcataaa taaggtgcga gaagatgtac
     1081 taaattcatt gaataacaac tttcttcaaa cgctaaatca agcttggaat gatcatcaaa
     1141 cagctatggt gatgattaga gacatactaa tgtacatgga ccgtgtgtat gtacaacaaa
     1201 ataatgtgga gaacgtctac aatttgggat taattatttt tcgagatcaa gttgtacgtt
     1261 atgggtgtat tagggatcat ctacggcaaa ctctattgga tatgattgca agagagcgga
     1321 aaggagaagt cgtagacaga ggcgcaataa gaaatgcttg ccagatgtta atgattttag
     1381 gtctcgaagg aagatcagtc tgtgaagaag attttgaggc tccttttttg gaaatgtctg
     1441 cagaattttt tcagatggaa agccagaaat ttttagcaga aaatagtgct tcagtatata
     1501 taaagaaagt agaagctaga attaatgaag aaatagaacg agtgatgcac tgccttgaca
     1561 aatcaacgga agaaccaatt gtaaaggtgg ttgaaaggga actcatttcc aagcacatga
     1621 agactatagt agaaatggag aattctgggc tagtacatat gttgaaaaat ggaaagacag
     1681 aagaccttgg ttgcatgtac aagttattta gtcgtgtgcc aaatggtttg aaaacaatgt
     1741 gtgagtgtat gagttcctat ttgagggagc aaggtaaagc tcttgtttct gaagaaggag
     1801 aaggaaagaa tcctgttgac tatatccagg gcttattgga tctgaagagt aggttcgatc
     1861 gcttcctcct ggaatcattc aacaatgacc gtctctttaa acaaactatt gcgggtgact
     1921 ttgagtattt tctcaacctc aactccaggt ctcctgaata cctctcatta tttattgatg
     1981 ataagctgaa aaagggagtc aaagggctaa cagaacaaga agtagaaaca atattggata
     2041 aagcaatggt cctttttagg tttatgcaag aaaaagatgt atttgaacgt tattataaac
     2101 aacacttggc aaggagactt ctcacaaata aaagtgtttc tgatgactct gaaaaaaaca
     2161 tgatatctaa gttaaagact gaatgtggat gtcagttcac gtcaaaactg gaaggaatgt
     2221 ttagggatat gagcatctca aacacaacga tggatgaatt caggcaacat ctacaggcaa
     2281 ctggtgtatc tttaggtggt gttgatctta cagtccgggt gctcacgaca ggatattggc
     2341 ccactcagtc agccacacca aagtgcaaca tcccaccagc accaagacat gcttttgaga
     2401 tattcagaag gttctactta gccaaacaca gtggtcgaca gctcacactc cagcatcata
     2461 tgggttctgc agatctcaat gccacatttt atggaccagt taaaaaggaa gatggatctg
     2521 aagttggtgt tggaggtgca caagtaactg gctctaatac acggaagcac atattgcaag
     2581 tttccacttt ccagatgacc atattaatgc tctttaataa tagagaaaaa tacacatttg
     2641 aggaaattca gcaagagaca gatatccctg aaagagagct tgttagagcc ctacagtccc
     2701 tcgcctgtgg taaaccaaca cagcgggttc ttacaaaaga acccaaatca aaggaaatag
     2761 aaaatggtca tatatttaca gttaatgatc aattcacatc caaactacac agagtcaaga
     2821 ttcaaacagt tgctgccaaa caaggtgaat ccgacccaga gaggaaagaa acaaggcaga
     2881 aagtagacga cgacagaaaa catgagatag aagctgctat agtgcggata atgaaatcta
     2941 gaaagaagat gcagcacaat gttctagtag cggaggtaac tcagcagttg aaggcgcgat
     3001 tcttaccaag tccagttgtt attaagaaac gtattgaagg acttattgag agagaatatt
     3061 tggcacgaac acctgaggat cgcaaagtat acacatatgt agcataaaat gcgttcagaa
     3121 atttgattta ttcttggact gtactcttcg catggactgg gaagttcttt taaatcatta
     3181 aatattaaga cgaccatctc ttctattaaa ttacagtaca tgttctagac cattgagatc
     3241 aagcctttac tccctttgag agtttccaac atcagttgat tgagcttcag gctttacaac
     3301 gtttatccct gtagagatca tctttacagt tcctcgggaa aatgtgaatg tgctgcgttt
     3361 tgttttcttt actgtatgaa aacaggaaaa taaaagagaa atttagaaaa tacagctcat
     3421 tacaataaaa ttgttggatt tcatttcaaa aaaaaaaaaa aaaaaactcg agcatgcatc
     3481 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     3541 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3601 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3661 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3721 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3781 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4081 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4141 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4201 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4261 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4321 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4381 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4441 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4501 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4561 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4621 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4681 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4741 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4801 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4861 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4921 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4981 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5041 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5101 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5161 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5221 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5281 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5341 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5401 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5461 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5521 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5581 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5641 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5701 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5761 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5821 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5881 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5941 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6001 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6061 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7261 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7561 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7921 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     7981 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     8041 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     8101 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     8161 ccagatatac gcgtt
//
