LOCUS       sequence-9696-ex        6041 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 6041)
  AUTHORS   Adams PD, Sellers WR, Sharma SK, Wu AD, Nalin CM, Kaelin WG
  TITLE     Identification of a cyclin-cdk2 recognition motif present in 
            substrates and p21-like cyclin-dependent kinase inhibitors.
  JOURNAL   Mol Cell Biol. 1996 Dec . 16(12):6623-33.
  PUBMED    8943316
REFERENCE   2  (bases 1 to 6041)
  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 
            Biol. 1996 Dec . 16(12):6623-33."
COMMENT     Sequence Label: p21-pRc/CMV
FEATURES             Location/Qualifiers
     source          1..6041
                     /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     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             926..1420
                     /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"
     primer_bind     complement(1472..1488)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1472..1488)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1498..1516)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1499..1516)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1536..1553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1542..1766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1812..2325
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1899..1918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2109..2130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2348..2368)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2353..2683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2534..2669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2750..3544
                     /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(2804..2823)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3414..3433
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3718..3852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3755..3774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3809..3828
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3884..3900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3884..3900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3897..3919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3908..3924
                     /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(3932..3938,3939..3956,3957..3962))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3977..3998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4115..4132)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4286..4874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4366..4385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5045..5836,5837..5905))
                     /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     5668..5687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5906..6010)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ctgcttaact ggcttatcga aattaatacg actcactata gggagaccca agcttgatat
      901 cgaattcgca ctcagaggag gcgccatgtc agaaccggct ggggatgtcc gtcagaaccc
      961 atgcggcagc aaggcctgcc gccgcctctt cggcccagtg gacagcgagc agctgagccg
     1021 cgactgtgat gcgctaatgg cgggctgcat ccaggaggcc cgtgagcgat ggaacttcga
     1081 ctttgtcacc gagacaccac tggagggtga cttcgcctgg gagcgtgtgc ggggccttgg
     1141 cctgcccaag ctctaccttc ccacggggcc ccggcgaggc cgggatgagt tgggaggagg
     1201 caggcggcct ggcacctcac ctgctctgct gcaggggaca gcagaggaag accatgtgga
     1261 cctgtcactg tcttgtaccc ttgtgcctcg ctcaggggag caggctgaag ggtccccagg
     1321 tggacctgga gactctcagg gtcgaaaacg gcggcagacc agcatgacag atttctacca
     1381 ctccaaacgc cggctgatct tctccaagag gaagccctaa ccgcccacag gaagcctgca
     1441 gtcctggagg gggggcccgg tacccccggg agatccacta gttctagagg gccctattct
     1501 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     1561 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1621 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1681 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     1741 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgag
     1801 gggggatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1861 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1921 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     1981 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2041 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2101 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2161 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     2221 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa tatttgctta
     2281 tacaatcttc ctgtttttgg ggcttttctg attatcaacc ggggtgggta ccgagctcga
     2341 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     2401 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2461 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2521 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2581 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2641 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2701 gatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2761 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2821 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2881 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     2941 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3001 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3061 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3121 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3181 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3241 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     3301 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3361 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3421 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3481 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3541 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3601 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3661 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     3721 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3781 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3841 catgtctgga tcccgtcgac ctcgagagct tggcgtaatc atggtcatag ctgtttcctg
     3901 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3961 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4021 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4081 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4141 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4201 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4261 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4321 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4381 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4441 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     4501 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4561 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4621 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4681 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4741 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4801 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4861 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4921 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4981 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5041 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5101 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5161 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5221 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5281 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5341 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5401 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5461 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5521 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5581 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5641 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5701 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5761 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5821 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5881 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5941 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6001 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
