LOCUS       sequence-134187-        7260 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 7260)
  AUTHORS   Ohta T, Xiong Y
  TITLE     Phosphorylation- and Skp1-independent in vitro ubiquitination of 
            E2F1 by multiple ROC-cullin ligases.
  JOURNAL   Cancer Res. 2001 Feb 15. 61(4):1347-53.
  PUBMED    11245432
REFERENCE   2  (bases 1 to 7260)
  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: "Cancer Res.
            2001 Feb 15. 61(4):1347-53."
COMMENT     Sequence Label: pcDNA3-myc3-bTrCP
FEATURES             Location/Qualifiers
     source          1..7260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        468..671
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     622..642
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     716..735
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        716..734
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             750..779
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             786..815
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             822..851
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2593..2658
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2666..2684)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2667..2684)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2704..2721)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2710..2934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2980..3408
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3067..3086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3277..3298
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3417..3437)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3422..3751
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3602..3737
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3664..3683
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3818..4612
                     /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(3872..3891)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4482..4501
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4786..4907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4823..4842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4877..4896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4956..4972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4956..4972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4969..4991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4980..4996
                     /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(5004..5010,5011..5028,5029..5034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5049..5070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5187..5204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5358..5946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5438..5457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6117..6908,6909..6977))
                     /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     6740..6759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6978..7082)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7157..7176)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
       61 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
      121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      661 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      721 cgactcacta tagggagacc caagctatgg agcaaaagct catctcagag gaagatctgg
      781 aattagagca aaagctcatc tcagaggaag atctggaatt agagcaaaag ctcatctcag
      841 aggaagatct ggaattgatg gacccggccg aggcggtgct gcaggagaaa gcgcttaagt
      901 ttatgaattc ctcagagaga gaagactgta ataatggcga accccctagg aagataatac
      961 cagagaagaa ttcacttaga cagacttaca acagctgtgc caggctttgc ataaaccaag
     1021 agacagtatg tctaacaagc actgctatga agactgaaaa ttgtgtggcc aaagccaaac
     1081 ttgccaatgg cacttccagc atgattgtgc ccaagcagcg gaaactctca gcaagctatg
     1141 agaaggaaaa ggagctgtgt gtcaagtatt ttgagcagtg gtcagagtct gatcaagtgg
     1201 aatttgtaga acaccttata tcccaaatgt gtcactacca gcatgggcac atcaactcct
     1261 acctaaaacc tatgctgcag agggatttca taactgcact gccagcacgg ggtctggacc
     1321 acatcgctga gaacattctg tcatacttgg acgccaagtc actgtgtgct gctgagctcg
     1381 tgtgcaagga atggtaccgc gtgacgtcgg acggcatgct gtggaaaaag ctcatcgaga
     1441 ggatggtcag gacggactct ctgtggcgag gcctggcaga gcgcagaggc tggggacagt
     1501 acttattcaa aaacaaacct cctgatgaga acgctcctcc caactccttt tatagagcgc
     1561 tttatcctaa aatcatacaa gacattgaga caatagagtc caattggaga tgtgggcgac
     1621 atagtttaca gagaatccac tgccggagtg aaacaagtaa aggggtttac tgtttacagt
     1681 acgacgacca gaagatagtc agcggccttc gagacaacac catcaagatc tgggataaaa
     1741 gcacactgga atgcaagcgg attctcacgg gccacacggg ctccgtcctg tgtctgcagt
     1801 acgatgagag ggtgatcatc acaggctcct cagactccac cgtcagagtg tgggatgtaa
     1861 atgcaggtga gatgctaaac acattgattc accactgtga agccgttctg cacctgcgct
     1921 tcaataatgg catgatggtg acctgttcca aagaccgttc catcgctgtg tgggatatgg
     1981 cttccccaac tgacatcacc ctcaggaggg tgctggtggg acaccgagct gcggtcaatg
     2041 ttgtagactt tgatgacaag tacatcgttt ctgcctctgg agatagaacc ataaaggtgt
     2101 ggaacacaag tacctgtgaa ttcgtaagga ccctaaatgg gcacaagcgt ggcatcgcct
     2161 gtttgcagta cagagacagg ctggtggtga gcggctcctc tgacaacacc atcaggctgt
     2221 gggacataga gtgtggagca tgcttgcgag tgttggaggg ccatgaggag ttggtacgct
     2281 gcattcgatt tgataacaaa aggatagtga gcggagccta tgatgggaaa attaaagtgt
     2341 gggatcttat ggctgctttg gacccgcgtg ctccagcagg gactctctgt ctgcggacac
     2401 ttgtggagca ttctggaaga gttttccgcc tccagtttga tgaattccag attgtcagta
     2461 gttcacatga tgacacaatt ctcatctggg acttcctgaa tgatccagct gctcacgctg
     2521 aaccgccccg ctccccttct cggatataca cctacatctc cagtacgtag tacgtaggct
     2581 agagttaaca tggactacaa agaccatgac ggtgattata aagatcatga tatcgattac
     2641 aaggatgacg atgacaagta gctattctat agtgtcacct aaatgctaga gctcgctgat
     2701 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2761 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2821 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2881 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2941 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3001 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3061 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3121 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3181 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3241 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3301 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3361 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3421 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3481 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3541 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3601 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3661 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3721 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3781 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3841 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3901 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3961 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4021 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4081 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4141 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4201 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4261 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4321 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4381 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4441 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4501 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4561 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4621 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4681 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4741 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4801 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4861 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4921 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4981 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5041 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5101 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5161 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5221 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5281 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5341 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5401 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5461 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5521 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5581 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5701 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5761 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5821 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5881 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5941 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6001 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6061 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6721 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6961 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7021 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7081 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7141 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7201 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
//
