LOCUS       sequence-232069-        7179 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 7179)
  AUTHORS   Hu J, Zacharek S, He YJ, Lee H, Shumway S, Duronio RJ, Xiong Y
  TITLE     WD40 protein FBW5 promotes ubiquitination of tumor suppressor TSC2 
            by DDB1-CUL4-ROC1 ligase.
  JOURNAL   Genes Dev. 2008 Apr 1. 22(7):866-71.
  PUBMED    18381890
REFERENCE   2  (bases 1 to 7179)
  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: "Genes Dev. 
            2008 Apr 1. 22(7):866-71."
COMMENT     Sequence Label: pCDNA3-myc3-FBW5
FEATURES             Location/Qualifiers
     source          1..7179
                     /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"
     promoter        complement(2734..2752)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2735..2752)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2772..2789)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2778..3002
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3048..3476
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3135..3154)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3345..3366
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3485..3505)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3490..3819
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3670..3805
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3732..3751
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3886..4680
                     /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(3940..3959)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4550..4569
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4854..4975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4891..4910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4945..4964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5024..5040)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5024..5040)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5037..5059)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5048..5064
                     /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(5072..5078,5079..5096,5097..5102))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5117..5138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5255..5272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5426..6014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5506..5525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6185..6976,6977..7045))
                     /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     6808..6827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7046..7150)
                     /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 gaattcacga tggacgaggg
     1021 cggcacgccc ctgctccccg acagcctggt ctaccagatc ttcctgagcc tgggcccggc
     1081 cgacgtgctg gccgccgggc tggtgtgccg ccaatggcag gccgtgtcgc gggacgagtt
     1141 cctgtggagg gagcagttct accgctacta ccaggtggcc cgcgacgtgc cccgacaccc
     1201 agcggccatg tcctggtacg aggagttcca gcggctgtat gacacggtgc cctgcgtgga
     1261 ggtgcagacg ctgcgggaac acacagacca ggtcctgcac ctcagcttct cccattccgg
     1321 gtaccagttc gcgtcctgct ccaaggactg cactgtgaag atctggagca acgacctgac
     1381 catctcgctg ctgcacagcg cggacatgcg gccctacaac tggagctaca cccagttctc
     1441 ccagttcaac aaggacgact cgctactgct ggcctcgggg gtgttcctgg ggccgcacaa
     1501 ctcctcatcc ggcgagattg ctgtcatcag cctagactcc ttcgcgctgc tgtcccgcgt
     1561 gcggaacaag ccctatgacg tgtttggctg ttggctcacc gagaccagcc tcatctcggg
     1621 gaacctgcac cgcatcggag atatcacctc ctgctcggtg ctgtggctca acaatgcctt
     1681 ccaggatgtg gagtcagaga acgtcaacgt ggtgaagcgg ctgttcaaga tccagaacct
     1741 caatgccagc accgtccgca cggtgatggt ggccgactgc agccgcttcg acagccctga
     1801 cctgctgctg gaagccggtg acccggccac gtccccctgc cgcatctttg acctgggcag
     1861 cgacaacgag gaggtggtgg ctggcccggc ccccgcccac gccaaggagg gcttgcggca
     1921 ctttctggat cgcgtgctgg aggggcgggc gcagccacag ctgtcggagc gcatgctaga
     1981 gaccaaggtg gccgagctgc tggcccaggg ccacaccaag ccacccgagc gcagtgccac
     2041 aggcgccaag agcaagtacc tcatcttcac cactggctgc ctcacctact ccccacacca
     2101 gatcggcatc aagcagatcc tgccacacca gatgaccacg gcagggcccg tgctgggtga
     2161 gggccggggc tccgatgcct tcttcgacgc gctggaccac gtcatagaca tacacggaca
     2221 catcatcggc atgggcctgt cgcccgacaa caggtacctg tacgtgaaca gccgcgcctg
     2281 gcccaacggt gcggtggtgg ccgaccccat gcagccgcca ccaatcgcgg aggagattga
     2341 cctgctggtg ttcgacctca agaccatgcg ggaggtgagg cgggctctgc gtgcgcaccg
     2401 cgcctacacg cccaacgacg agtgcttctt catcttcctg gacgtcagca gggacttcgt
     2461 ggccagcggg gcggaggacc ggcacggcta catctgggac cgccactaca acatctgtct
     2521 ggccaggctg cggcacgagg atgtggtcaa ctcagtggtc ttcagtcccc aggagcagga
     2581 gctgctgctc acggccagcg acgacgccac catcaaagcc tggcgctccc cacgcaccat
     2641 gcgcgtcctc caggcacctc gcccacggcc tcgcaccttc ttctcctggc ttgccagcca
     2701 gaggcgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     2761 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2821 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2881 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2941 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3001 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3061 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3121 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3181 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3241 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3301 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3361 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3421 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3481 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3541 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3661 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3721 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3781 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3841 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3901 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3961 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4021 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4081 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4141 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4201 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4261 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4321 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4381 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4441 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4501 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4561 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4621 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4681 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4741 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4801 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4981 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5041 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5101 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5161 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5221 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5281 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5341 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5401 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5461 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5521 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5581 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5641 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5701 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5761 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5821 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5881 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5941 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6001 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6061 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6121 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6181 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7081 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7141 gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
