LOCUS       sequence-429310-        5232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressesp21(dPCNA)-mCerulean in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5232)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 5232)
  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: "Mol Cell. 
            2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: p21(dPCNA)-mCerulean-NeoR
FEATURES             Location/Qualifiers
     source          1..5232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             604..1095
                     /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
                     TAEEDHVDLSLSCTLVPRSGEQAEGSPGGPGDSQGRKRRQTSATAAYHSKRRLIFSKRK
                     P"
     regulatory      1108..1117
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1114..1116,1117..1119,1120..1830)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1159..1180)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1420..1439)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1767..1788
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1831..1896
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2020..2141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2057..2076)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2111..2130
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2148..2603)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2285..2306)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2497..2516
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2630..2734
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2736..3093
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2736..2927
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2759..2779)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2944..3079
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3128..3922
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3128..3922
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3182..3201)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3792..3811
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4110..4129)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4154..4201
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4530..5118
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5019..5038
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccgcc
      601 accatgtcag aaccggctgg ggatgtccgt cagaacccat gcggcagcaa ggcctgccgc
      661 cgcctcttcg gcccagtgga cagcgagcag ctgagccgcg actgtgatgc gctaatggcg
      721 ggctgcatcc aggaggcccg tgagcgatgg aacttcgact ttgtcaccga gacaccactg
      781 gagggtgact tcgcctggga gcgtgtgcgg ggccttggcc tgcccaagct ctaccttccc
      841 acggggcccc ggcgaggccg ggatgagttg ggaggaggca ggcggcctgg cacctcacct
      901 gctctgctgc aggggacagc agaggaagac catgtggacc tgtcactgtc ttgtaccctt
      961 gtgcctcgct caggggagca ggctgaaggg tccccaggtg gacctggaga ctctcagggt
     1021 cgaaaacggc ggcagaccag cgcgacagct gcctaccact ccaaacgccg gctgatcttc
     1081 tccaagagga agcccggagg accggtcgcc accatggtga gcaagggcga ggagctgttc
     1141 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1201 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1261 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg
     1321 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1381 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1441 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1501 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacgc catcagcgac
     1561 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1621 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1681 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
     1741 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1801 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg
     1861 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata
     1921 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1981 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2041 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2101 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2161 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2221 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2281 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2341 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2401 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2461 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2521 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2581 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2641 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2701 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     2761 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2821 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2941 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3001 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3061 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3121 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3181 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3241 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3301 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3361 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3421 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3481 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3541 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3601 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     3661 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3721 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3781 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3841 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3901 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3961 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4021 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4081 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4141 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4201 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4261 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4321 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4381 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4441 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4501 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4561 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4621 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4681 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4741 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4801 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4861 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4921 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4981 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5041 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5101 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5161 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5221 accgccatgc at
//
