LOCUS       sequence-190558-        5297 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 5297)
  AUTHORS   Rajagopalan H, Jallepalli PV, Rago C, Velculescu VE, Kinzler KW, 
            Vogelstein B, Lengauer C
  TITLE     Inactivation of hCDC4 can cause chromosomal instability.
  JOURNAL   Nature. 2004 Mar 4. 428(6978):77-81.
  PUBMED    14999283
REFERENCE   2  (bases 1 to 5297)
  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: "Nature. 
            2004 Mar 4. 428(6978):77-81."
COMMENT     Sequence Label: pCMV-Myc CDC4 WT*
FEATURES             Location/Qualifiers
     source          1..5297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          646..742
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     polyA_signal    2410..2544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2434..2453)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2488..2507
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2655..2671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2655..2671)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2668..2690)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2679..2695
                     /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(2703..2709,2710..2727,2728..2733))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2748..2769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2886..2903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3057..3645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3137..3156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3816..4607,4608..4676))
                     /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     4439..4458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4677..4781)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4849..4867
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4905..4927)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5027..5046
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5240..5262
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5254..5271
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5255..5271
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaaaaac tgctcctcag
      721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      781 gaattgtacc cgcgggccca ccatggcatc aatgcagaag ctgatctcag aggaggacct
      841 gcttatggcc atggaggccc gaattcggtc gaccgagatc ttaccatgtt cagcaacacc
      901 aacaactttt ggggacctca gagcagccaa tggccaaggg caacaacgac gccgaattac
      961 atctgtccag ccacctacag gcctccagga atggctaaaa atgtttcaga gctggagtgg
     1021 accagagaaa ttgcttgctt tagatgaact cattgatagt tgtgaaccaa cacaagtaaa
     1081 acatatgatg caagtgatag aaccccagtt tcaacgagac ttcatttcat tgctccctaa
     1141 agagttggca ctctatgtgc tttcattcct ggaacccaaa gacctgctac aagcagctca
     1201 gacatgtcgc tactggagaa ttttggctga agacaacctt ctctggagag agaaatgcaa
     1261 agaagagggg attgatgaac cattgcacat caagagaaga aaagtaataa aaccaggttt
     1321 catacacagt ccatggaaaa gtgcatacat cagacagcac agaattgata ctaactggag
     1381 gcgaggagaa ctcaaatctc ctaaggtgct gaaaggacat gatgatcatg tgatcacatg
     1441 cttacagttt tgtggtaacc gaatagttag tggttctgat gacaacactt taaaagtttg
     1501 gtcagcagtc acaggcaaat gtctgagaac attagtggga catacaggtg gagtatggtc
     1561 atcacaaatg agagacaaca tcatcattag tggatctaca gatcggacac tcaaagtgtg
     1621 gaatgcagag actggagaat gtatacacac cttatatggg catacttcca ctgtgcgttg
     1681 tatgcatctt catgaaaaaa gagttgttag cggttctcga gatgccactc ttagggtttg
     1741 ggatattgag acaggccagt gtttacatgt tttgatgggt catgttgcag cagtccgctg
     1801 tgttcaatat gatggcagga gggttgttag tggagcatat gattttatgg taaaggtgtg
     1861 ggatccagag gctgaaacct gtctacacac gttgcagggg catactaata gagtctattc
     1921 attacagttt gatggtatcc atgtggtgag tggatctctt gatacatcaa tccgtgtttg
     1981 ggatgtggag acagggaatt gcattcacac gttaacaggg caccagtcgt taacaagtgg
     2041 aatggaactc aaagacaata ttcttgtttc tgggaatgca gattctacag ttaaaatctg
     2101 ggatatcaaa acaggacagt gtgtacaaac attgcaaggt cccaacaagc atcagagtgc
     2161 tgtgacctgt ttacagttca acaagaactt tgtaactacc agctcagatg atggaactgt
     2221 agaactatgg gacttgaaaa cgggtgaatt tattcgaaac ctagtcacat tggagagtgg
     2281 ggggagtggg ggagttgtgt ggcggatcag agcctcaaac acaaagctgg tgtgtgcagt
     2341 tgggagtcgg aatgggactg aagaaaccaa gctgctggtg ctggactttg atgtggacgc
     2401 ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2461 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2521 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2581 cagggggagg tgtgggaggt tttttcggat cctctagagt cgatctgcag gcatgctagc
     2641 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2701 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2761 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2821 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4741 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4801 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4861 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4921 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4981 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     5041 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5101 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5161 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     5221 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagttcgag
     5281 cttgcatgcc tgcaggt
//
