LOCUS       sequence-427411-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses p21-HA(d17-24) in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  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 5867)
  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-HA(d17-24)
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1363..1389
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1420..1438)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1421..1438)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1458..1475)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1464..1688
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1734..2162
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1821..1840)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2031..2052
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2171..2191)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2176..2505
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2356..2491
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2418..2437
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2572..3366
                     /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(2626..2645)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3236..3255
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3540..3661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3577..3596)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3631..3650
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3710..3726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3710..3726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3723..3745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3734..3750
                     /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(3758..3764,3765..3782,3783..3788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3803..3824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3941..3958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4112..4700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4192..4211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4871..5662,5663..5731))
                     /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     5494..5513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5732..5836)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatgtca
      901 gaaccggctg gggatgtccg tcagaaccca tgcggcagca aggtggacag cgagcagctg
      961 agccgcgact gtgatgcgct aatggcgggc tgcatccagg aggcccgtga gcgatggaac
     1021 ttcgactttg tcaccgagac accactggag ggtgacttcg cctgggagcg tgtgcggggc
     1081 cttggcctgc ccaagctcta ccttcccacg gggccccggc gaggccggga tgagttggga
     1141 ggaggcaggc ggcctggcac ctcacctgct ctgctgcagg ggacagcaga ggaagaccat
     1201 gtggacctgt cactgtcttg tacccttgtg cctcgctcag gggagcaggc tgaagggtcc
     1261 ccaggtggac ctggagactc tcagggtcga aaacggcggc agaccagcat gacagatttc
     1321 taccactcca aacgccggct gatcttctcc aagaggaagc cctacccata cgacgtccca
     1381 gactacgctt aactcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     1441 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1501 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1561 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1621 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     1681 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     1741 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1801 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1861 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1921 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1981 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2041 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2101 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2161 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2221 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2281 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2341 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2401 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     2461 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     2521 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     2581 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2641 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     2701 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     2761 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     2821 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     2881 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     2941 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3001 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3061 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3121 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3181 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3241 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3301 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3361 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3421 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3481 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3541 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3601 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3661 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3721 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3781 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3841 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3901 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3961 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4921 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4981 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5041 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5101 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5161 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5221 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5281 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5341 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5401 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5461 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5521 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5581 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5641 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5701 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5761 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5821 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
