LOCUS       sequence-191933-        5301 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 5301)
  AUTHORS   Luo Y, Hurwitz J, Massague J
  TITLE     Cell-cycle inhibition by independent CDK and PCNA binding domains in
            p21Cip1.
  JOURNAL   Nature. 1995 May 11. 375(6527):159-61.
  PUBMED    7753174
REFERENCE   2  (bases 1 to 5301)
  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. 
            1995 May 11. 375(6527):159-61."
COMMENT     Sequence Label: pCMV5 human p27
FEATURES             Location/Qualifiers
     source          1..5301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        38..417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        418..621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     572..592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1362..1984
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1561..1580)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2008..2028)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2013..2342
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2193..2328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2380..2399)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2381..2399)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2413..2429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2413..2429)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2426..2448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2437..2453
                     /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(2461..2467,2468..2485,2486..2491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2506..2527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2644..2661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2815..3403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2895..2914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3574..4365,4366..4434))
                     /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     4197..4216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4435..4539)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4566..5021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4653..4672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4863..4884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5147..5169
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5161..5178
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5162..5178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat
       61 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      121 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      181 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      241 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
      301 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      361 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      421 atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca
      481 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      541 ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg
      601 gaggtctata taagcagagc tcgtttagtg aaccgtcaga attcagatct ggtaccgccg
      661 cgtaggggcg ctttgttttg ttcggttttg tttttttgag agagcgagag aggcggtcgt
      721 gcagacccgg gagaaagctc ggctagcatg tcaaacgtgc gagtgtctaa cgggagccct
      781 agcctggagc ggatggacgc caggcaggcg gagcacccca agccctcggc ctgcaggaac
      841 ctcttcggcc cggtggacca cgaagagtta acccgggact tggagaagca ctgcagagac
      901 atggaagagg cgagccagcg caagtggaat ttcgattttc agaatcacaa acccctagag
      961 ggcaagtacg agtggcaaga ggtggagaag ggcagcttgc ccgagttcta ctacagaccc
     1021 ccgcggcccc ccaaaggtgc ctgcaaggtg ccggcgcagg agagccagga tgtcagcggg
     1081 agccgcccgg cggcgccttt aattggggct ccggctaact ctgaggacac gcatttggtg
     1141 gacccaaaga ctgatccgtc ggacagccag acggggttag cggagcaatg cgcaggaata
     1201 aggaagcgac ctgcaaccga cgattcttct actcaaaaca aaagagccaa cagaacagaa
     1261 gaaaatgttt cagacggttc cccaaatgcc ggttctgtgg agcagacgcc caagaagcct
     1321 ggcctcagaa gacgtcaaac gtaacggagc ttaaggatcc cgggtggcat ccctgtgacc
     1381 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     1441 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     1501 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     1561 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     1621 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     1681 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     1741 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     1801 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     1861 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     1921 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     1981 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2041 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2101 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2161 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2221 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2281 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2341 aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa
     2401 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2461 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2521 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2581 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2641 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2701 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2761 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2821 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2881 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2941 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3001 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3061 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3121 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3181 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3241 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3301 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3421 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3481 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3541 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3601 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3661 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3721 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3781 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3841 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3901 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3961 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4021 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4081 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4141 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4201 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4261 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4321 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4381 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4441 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4501 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4561 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4621 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4681 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4741 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4801 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4861 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4921 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4981 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
     5041 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     5101 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     5161 tgtaaaacga cggccagtgc caagctgatc tatacattga atcaatattg gcaattagcc
     5221 atattagtca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
     5281 tatctatatc ataatatgta c
//
