LOCUS       sequence-214193-        8264 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 8264)
  AUTHORS   Xu Y, Sengupta T, Kukreja L, Minella AC
  TITLE     MicroRNA-223 regulates cyclin E activity by modulating expression of
            F-box and WD-40 domain protein 7.
  JOURNAL   J Biol Chem. 2010 Nov 5;285(45):34439-46. doi: 
            10.1074/jbc.M110.152306. Epub 2010 Sep 7.
  PUBMED    20826802
REFERENCE   2  (bases 1 to 8264)
  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"; doi: 
            "10.1074/jbc.M110.152306"; journalName: "J Biol Chem"; date: 
            "2010-11-5- 5"; volume: "285"; issue: "45"; pages: "34439-46"
COMMENT     Sequence Label: pFLAG-Fbw7-alpha-3'UTR
FEATURES             Location/Qualifiers
     source          1..8264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4569..4598
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    4605..5227
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4804..4823)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5251..5271)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5256..5585
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5436..5571
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5623..5642)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5624..5642)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5656..5672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5656..5672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5669..5691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5680..5696
                     /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(5704..5710,5711..5728,5729..5734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5749..5770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5887..5904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6058..6646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6138..6157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6817..7608,7609..7677))
                     /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     7440..7459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7678..7782)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7809..8237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7896..7915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8106..8127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgaatcagga actgctctct
     1021 gtgggcagca aaagacgacg aactggaggc tctctgagag gtaacccttc ctcaagccag
     1081 gtagatgaag aacagatgaa tcgtgtggta gaggaggaac agcaacagca actcagacaa
     1141 caagaggagg agcacactgc aaggaatggt gaagttgttg gagtagaacc tagacctgga
     1201 ggccaaaatg attcccagca aggacagttg gaagaaaaca ataatagatt tatttcggta
     1261 gatgaggact cctcaggaaa ccaagaagaa caagaggaag atgaagaaca tgctggtgaa
     1321 caagatgagg aggatgagga ggaggaggag atggaccagg agagtgacga ttttgatcag
     1381 tctgatgata gtagcagaga agatgaacat acacatacta acagtgtcac gaactccagt
     1441 agtattgtgg acctgcccgt tcaccaactc tcctccccat tctatacaaa aacaacaaaa
     1501 atgaaaagaa agttggacca tggttctgag gtccgctctt tttctttggg aaagaaacca
     1561 tgcaaagtct cagaatatac aagtaccact gggcttgtac catgttcagc aacaccaaca
     1621 acttttgggg acctcagagc agccaatggc caagggcaac aacgacgccg aattacatct
     1681 gtccagccac ctacaggcct ccaggaatgg ctaaaaatgt ttcagagctg gagtggacca
     1741 gagaaattgc ttgctttaga tgaactcatt gatagttgtg aaccaacaca agtaaaacat
     1801 atgatgcaag tgatagaacc ccagtttcaa cgagacttca tttcattgct ccctaaagag
     1861 ttggcactct atgtgctttc attcctggaa cccaaagacc tgctacaagc agctcagaca
     1921 tgtcgctact ggagaatttt ggctgaagac aaccttctct ggagagagaa atgcaaagaa
     1981 gaggggattg atgaaccatt gcacatcaag agaagaaaag taataaaacc aggtttcata
     2041 cacagtccat ggaaaagtgc atacatcaga cagcacagaa ttgatactaa ctggaggcga
     2101 ggagaactca aatctcctaa ggtgctgaaa ggacatgatg atcatgtgat cacatgctta
     2161 cagttttgtg gtaaccgaat agttagtggt tctgatgaca acactttaaa agtttggtca
     2221 gcagtcacag gcaaatgtct gagaacatta gtgggacata caggtggagt atggtcatca
     2281 caaatgagag acaacatcat cattagtgga tctacagatc ggacactcaa agtgtggaat
     2341 gcagagactg gagaatgtat acacacctta tatgggcata cttccactgt gcgttgtatg
     2401 catcttcatg aaaaaagagt tgttagcggt tctcgagatg ccactcttag ggtttgggat
     2461 attgagacag gccagtgttt acatgttttg atgggtcatg ttgcagcagt ccgctgtgtt
     2521 caatatgatg gcaggagggt tgttagtgga gcatatgatt ttatggtaaa ggtgtgggat
     2581 ccagagactg aaacctgtct acacacgttg caggggcata ctaatagagt ctattcatta
     2641 cagtttgatg gtatccatgt ggtgagtgga tctcttgata catcaatccg tgtttgggat
     2701 gtggagacag ggaattgcat tcacacgtta acagggcacc agtcgttaac aagtggaatg
     2761 gaactcaaag acaatattct tgtctctggg aatgcagatt ctacagttaa aatctgggat
     2821 atcaaaacag gacagtgttt acaaacattg caaggtccca acaagcatca gagtgctgtg
     2881 acctgtttac agttcaacaa gaactttgta attaccagct cagatgatgg aactgtaaaa
     2941 ctatgggact tgaaaacggg tgaatttatt cgaaacctag tcacattgga gagtgggggg
     3001 agtgggggag ttgtgtggcg gatcagagcc tcaaacacaa agctggtgtg tgcagttggg
     3061 agtcggaatg ggactgaaga aaccaagctg ctggtgctgg actttgatgt ggaatgaagt
     3121 gagaattgtg tagacgatat actccctgcc cttccccctg caaaaagaaa aaaagaaaag
     3181 aaaaagaaaa aaatcccttg ttctcagtgg tgcaggatgt tggcttgggg caacagattg
     3241 aaaagaccta cagactaaga aggaaaagaa gaagagatga caaaccataa ctgacaagag
     3301 aggcgtctgc tgtctcatca cataaaaggc ttcactcttg actgagggca gctttgcaaa
     3361 atgagacttt ctaaaacaaa ccaggtgcaa ttatttcttt attttcttct ccagtggtca
     3421 ttgggcagtg ttaatgctga aacatcatta cagattctgc tagcctgttc ttttaccact
     3481 gacagctaga cacctagaaa ggaactgcaa taatatcaaa acaagtactg gttgactttc
     3541 taattagaga gcatctgcaa caaaaagtca tttttcagga gtggaaaagc ttaaaaaaat
     3601 tactgtgaat tgtttttgta cagttatcat gaaaagcttt tttttttttc ttcttgccaa
     3661 ccattgccaa tgtcaatcaa tcacagtatt agcctctgtt aatctattta ctgttgcttc
     3721 catatacatt cttcaatgca tatgttgctc aaaggtggca agttgtcctg ggttctgtga
     3781 gtcctgagat ggatttaatt cttgatgctg gtgctggaag taggtcttca aatatgggat
     3841 tgttgtccca accctgtact gtactcccag tggccaaact tatttatgct gctaaatgaa
     3901 agaaagaaaa aagcaaatta ttttttttta tttttttttc tgctgtgacg ttttagtccc
     3961 agactgaatt ccaaatttgc tctagtttgg ttatggaaaa aagacttttt gccactgaaa
     4021 cttgagccat ctgtgcctct aagaggctga gaatggaaga gtttcagata ataaagagtg
     4081 aagtttgcct gcaagtaaag aattgagagt gtgtgcaaag cttattttct tttatctggg
     4141 caaaaattaa aacacattcc ttggaacaga gctattactt gcctgttctg tggagaaact
     4201 tttctttttg agggctgtgg tgaatggatg aacgtacatc gtaaaactga caaaatattt
     4261 taaaaatata taaaacacaa aattaaaata aagttgctgg tcagtcttag tgttttacag
     4321 tatttgggaa aacaactgtt acagttttat tgctctgagt aactgacaaa gcagaaacta
     4381 ttcagttttt gtagtaaagg cgtcacatgc aaacaaacaa aatgaatgaa acagtcaaat
     4441 ggtttgcctc attctccaag agccacaact caagctgaac tgtgaaagtg gtttaacact
     4501 agatctgata tcggtaccag tcgactctag aggatctgat atcggtacca gtcgactcta
     4561 gaggatccga acaaaaactc atctcagaag aggatctgtg acccgggtgg catccctgtg
     4621 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     4681 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     4741 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     4801 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     4861 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     4921 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     4981 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     5041 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat
     5101 accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac
     5161 atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg
     5221 ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5281 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5341 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5401 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5461 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5521 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5581 gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat
     5641 taaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5701 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5761 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5821 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5881 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5941 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgtgaagcgt
     6241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6901 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6961 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7021 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7081 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7141 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7201 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7261 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7321 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7381 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7441 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7501 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7561 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7621 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7681 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7801 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     7861 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     7921 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     7981 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     8041 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     8101 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     8161 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     8221 aaaatttaac gcgaatttta acaaaatatt aacgcttaca attt
//
