LOCUS       sequence-190134-        7891 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 7891)
  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 7891)
  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-CUL5
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             899..928
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             935..964
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             971..1000
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3369..3434
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3446..3464)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3447..3464)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3484..3501)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3490..3714
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3760..4188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3847..3866)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4057..4078
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4197..4217)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4202..4531
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4382..4517
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4444..4463
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4598..5392
                     /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(4652..4671)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5262..5281
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5566..5687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5603..5622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5657..5676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5736..5752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5736..5752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5749..5771)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5760..5776
                     /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(5784..5790,5791..5808,5809..5814))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5829..5850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5967..5984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6138..6726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6218..6237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6897..7688,7689..7757))
                     /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     7520..7539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7758..7862)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctatgga
      901 gcaaaagctc atctcagagg aagatctgga attagagcaa aagctcatct cagaggaaga
      961 tctggaatta gagcaaaagc tcatctcaga ggaagatctg gaattcagga tcccggcgac
     1021 gtctaatctg ttaaagaata aaggttctct tcagtttgaa gacaaatggg attttatgcg
     1081 cccgattgtt ttgaagcttt tacgccagga atctgttaca aaacagcagt ggtttgatct
     1141 gttttcggat gtgcatgcag tctgtctttg ggatgataaa ggcccagcaa aaattcatca
     1201 ggctttaaaa gaagatattc ttgagtttat taagcaggca caggcacgag tactgagcca
     1261 tcaagatgat acggctttgc taaaagcata tattgttgaa tggcgaaagt tctttacaca
     1321 atgtgatatt ttaccaaaac ctttttgtca actagagatt actttaatgg gtaaacaggg
     1381 cagcaataaa aaatcaaatg tggaagacag tattgttcga aagcttatgc ttgatacatg
     1441 gaatgagtca atcttttcaa acataaaaaa cagactccaa gatagtgcaa tgaagctggt
     1501 acatgctgag agattgggag aagcttttga ttctcagctg gttattggag taagagaatc
     1561 ctatgttaac ctttgttcta atcctgagga taaacttcaa atttataggg acaattttga
     1621 gaaggcatac ttggattcaa cagagagatt ttatagaaca caagcaccct cgtatttaca
     1681 acaaaatggt gtacagaatt atatgaaata tgcagatgct aaattaaaag aagaagaaaa
     1741 acgagcacta cgttatttag aaacaagacg agaatgtaac tccgttgaag cactcatgga
     1801 atgctgtgta aatgccctgg tgacatcatt taaagagact atcttagctg agtgccaagg
     1861 catgatcaag agaaatgaaa ctgaaaaatt acatttaatg ttttcattga tggacaaagt
     1921 tcctaatggt atagagccaa tgttgaaaga cttggaggaa catatcatta gtgctggcct
     1981 ggcagatatg gtagcagctg ctgaaactat tactactgac tctgagaaat acgttgagca
     2041 gttacttaca ctatttaata gatttagtaa actcgtcaaa gaagcttttc aagatgatcc
     2101 acgatttctt actgcaagag ataaggcgta taaagcagtt gttaatgatg ctaccatatt
     2161 taaacttgaa ttacctttga agcagaaggg ggtgggatta aaaactcagc ctgaatcaaa
     2221 atgccctgag ctgcttgcca attactgtga catgttgcta agaaaaacac cattaagcaa
     2281 aaaactaacc tctgaagaga ttgaagcaaa gcttaaagaa gtgctcttgg tacttaagta
     2341 tgtacagaac aaagatgttt ttatgaggta tcataaagct catttgacac gacgtcttat
     2401 attagacatc tctgccgata gtgaaattga agaaaacatg gtagagtggc taagagaagt
     2461 tggtatgcca gcggattatg taaacaagct tgctagaatg tttcaggaca taaaagtatc
     2521 tgaagatttg aaccaagctt ttaaggaaat gcacaaaaat aataaattgg cattaccagc
     2581 tgattcagtt aatataaaaa ttctgaatgc tggcgcctgg tcaagaagtt ctgagaaagt
     2641 ctttgtctca cttcctactg aactggagga cttgataccg gaagtagaag aattctacaa
     2701 aaaaaatcat agtggtagaa aattacattg gcatcatctc atgtcaaatg gaattataac
     2761 atttaagaat gaagttggtc aatatgattt ggaggtaacc acgtttcagc tcgctgtatt
     2821 gtttgcatgg aaccaaagac ccagagagaa aatcagcttt gaaaatctta agcttgcaac
     2881 tgaactccct gatgctgaac ttaggaggac tttatggtct ttagtagctt tcccaaaact
     2941 caaacggcaa gttttgttgt atgaacctca agtcaactca cccaaagact ttacagaagg
     3001 taccctcttc tcagtgaacc aggagttcag tttaataaaa aatgcaaagg ttcagaaaag
     3061 gggtaaaatc aacttgattg gacgtttgca gctcactaca gaaaggatga gagaagaaga
     3121 gaatgaagga atagttcaac tacgaatact aagaacccag gaagctatca tacaaataat
     3181 gaaaatgaga aagaaaatta gtaatgctca gctgcagact gaattagtag aaattttgaa
     3241 aaacatgttc ttgccacaaa agaaaatgat aaaagagcaa atagagtggc taatagagca
     3301 caaatacatc agaagagatg aatctgatat caacactttc atatatatgg catctagagt
     3361 taaccatgga ctacaaagac catgacggtg attataaaga tcatgatatc gattacaagg
     3421 atgacgatga caagtagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     3481 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3541 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3601 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3661 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3721 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3781 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3841 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3901 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3961 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4021 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4081 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4141 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4201 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4261 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4321 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4381 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4441 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4501 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4561 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4621 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4681 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4741 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4801 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4861 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4921 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4981 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5041 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5101 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5161 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5221 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5281 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5341 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5401 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5461 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5521 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5581 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5641 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5701 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5761 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5821 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5881 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5941 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6001 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6061 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6121 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6181 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6241 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6301 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6361 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6421 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6481 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6541 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6601 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6661 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6721 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6781 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6841 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6901 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6961 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7021 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7081 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7141 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7201 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7261 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7321 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7381 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7441 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7501 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7561 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7621 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7681 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7741 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7801 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7861 cgcacatttc cccgaaaagt gccacctgac g
//
