LOCUS       sequence-137773-        5223 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 5223)
  AUTHORS   Zhou BP, Liao Y, Xia W, Spohn B, Lee MH, Hung MC
  TITLE     Cytoplasmic localization of p21Cip1/WAF1 by Akt-induced 
            phosphorylation in HER-2/neu-overexpressing cells.
  JOURNAL   Nat Cell Biol. 2001 Mar . 3(3):245-52.
  PUBMED    11231573
REFERENCE   2  (bases 1 to 5223)
  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: "Nat Cell 
            Biol. 2001 Mar . 3(3):245-52."
COMMENT     Sequence Label: Flag p21 WT
FEATURES             Location/Qualifiers
     source          1..5223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     RBS             668..690
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             700..723
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             745..1239
                     /codon_start=1
                     /gene="Homo sapiens CDKN1A"
                     /product="human cyclin-dependent kinase inhibitor 1A"
                     /label=p21
                     /note="mediates p53-dependent arrest in the G1 phase of the
                     cell cycle"
                     /translation="MSEPAGDVRQNPCGSKACRRLFGPVDSEQLSRDCDALMAGCIQEA
                     RERWNFDFVTETPLEGDFAWERVRGLGLPKLYLPTGPRRGRDELGGGRRPGTSPALLQG
                     TAEEDHVDLSLSCTLVPRSGEQAEGSPGGPGDSQGRKRRQTSMTDFYHSKRRLIFSKRK
                     P"
     polyA_signal    1247..1869
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1446..1465)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1893..1913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1898..2227
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2078..2213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2265..2284)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2266..2284)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2298..2314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2298..2314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2311..2333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2322..2338
                     /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(2346..2352,2353..2370,2371..2376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2391..2412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2529..2546)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2700..3288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2780..2799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3459..4250,4251..4319))
                     /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     4082..4101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4320..4424)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4451..4906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4538..4557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4748..4769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5032..5054
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5046..5063
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5047..5063
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattcaga tctggtacca cgcgtatcga taagcttgca tgcctgcagg tcgactctag
      661 aaataatttt gtttaacttt aagaaggaga tataccatgg actacaaaga cgatgacgat
      721 aaagcaagaa gagcatctgt gcatatgtca gaaccggctg gggatgtccg tcagaaccca
      781 tgcggcagca aggcctgccg ccgcctcttc ggcccagtgg acagcgagca gctgagccgc
      841 gactgtgatg cgctaatggc gggctgcatc caggaggccc gtgagcgatg gaacttcgac
      901 tttgtcaccg agacaccact ggagggtgac ttcgcctggg agcgtgtgcg gggccttggc
      961 ctgcccaagc tctaccttcc cacggggccc cggcgaggcc gggatgagtt gggaggaggc
     1021 aggcggcctg gcacctcacc tgctctgctg caggggacag cagaggaaga ccatgtggac
     1081 ctgtcactgt cttgtaccct tgtgcctcgc tcaggggagc aggctgaagg gtccccaggt
     1141 ggacctggag actctcaggg tcgaaaacgg cggcagacca gcatgacaga tttctaccac
     1201 tccaaacgcc ggctgatctt ctccaagagg aagccctaag gatcccgggt ggcatccctg
     1261 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     1321 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     1381 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     1441 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     1501 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     1561 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     1621 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     1681 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     1741 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     1801 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     1861 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     1921 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1981 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2041 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2101 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2161 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2221 ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat agtgagtcgt
     2281 attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2341 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2401 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2461 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2521 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2581 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2641 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2701 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2761 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2821 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2881 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2941 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3001 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3061 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3121 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3181 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3241 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3301 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3361 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3421 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3481 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3541 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3601 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3661 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3721 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3781 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3841 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3901 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3961 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4021 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4081 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4141 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4201 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4261 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4321 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4381 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4441 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4501 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4561 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4621 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4681 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4741 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4801 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4861 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca
     4921 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     4981 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     5041 gacgttgtaa aacgacggcc agtgccaagc tgatctatac attgaatcaa tattggcaat
     5101 tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
     5161 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
     5221 gtt
//
