LOCUS       sequence-322389-        7911 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 7911)
  AUTHORS   Popov N, Wanzel M, Madiredjo M, Zhang D, Beijersbergen R, Bernards 
            R, Moll R, Elledge SJ, Eilers M
  TITLE     The ubiquitin-specific protease USP28 is required for MYC stability.
  JOURNAL   Nat Cell Biol. 2007 Jul . 9(7):765-74.
  PUBMED    17558397
REFERENCE   2  (bases 1 to 7911)
  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. 2007 Jul . 9(7):765-74."
COMMENT     Sequence Label: pDZ Flag USP28
FEATURES             Location/Qualifiers
     source          1..7911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(562..1353,1354..1422))
                     /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     1185..1204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1423..1527)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1554..2009
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1641..1660)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1851..1872
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2135..2157
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2149..2166
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2150..2166
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2327..2706
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2707..2910
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2861..2881
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2944..2967
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3004..3028
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6203..6227)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6261..6883
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6460..6479)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6907..6927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6912..7241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7092..7227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7279..7298)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7280..7298)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7312..7328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7312..7328)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7325..7347)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7336..7352
                     /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(7360..7366,7367..7384,7385..7390))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7405..7426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7543..7560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7714..7911,1..391))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7794..7813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
       61 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      121 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      181 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      241 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      301 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      361 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
      421 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      481 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
      541 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
      601 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
      661 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
      721 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
      781 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      841 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
      901 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      961 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1021 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1081 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1141 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1201 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1261 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1321 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1381 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1441 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1501 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     1561 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1621 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1681 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1741 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1801 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1861 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1921 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1981 ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg
     2041 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     2101 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     2161 gccagtgcca agctgatcta tacattgaat caatattggc aattagccat attagtcatt
     2221 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     2281 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg
     2341 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     2401 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     2461 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     2521 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     2581 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     2641 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     2701 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     2761 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     2821 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
     2881 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagaa ttgatctacc
     2941 atggactaca aagacgatga cgacaagctt gcggccgcga attcatcgat agatctgata
     3001 tcaacaagtt tgtacaaaaa agcaggctcc gcggccgccc ccttcaccat gactgcggag
     3061 ctgcagcagg acgacgcggc cggcgcggca gacggccacg gctcgagctg ccaaatgctg
     3121 ttaaatcaac tgagagaaat cacaggcatt caggaccctt cctttctcca tgaagctctg
     3181 aaggccagta atggtgacat tactcaggca gtcagccttc tcactgatga gagagttaag
     3241 gagcccagtc aagacactgt tgctacagaa ccatctgaag tagaggggag tgctgccaac
     3301 aaggaagtat tagcaaaagt tatagacctt actcatgata acaaagatga tcttcaggct
     3361 gccattgctt tgagtctact ggagtctccc aaaattcaag ctgatggaag agatcttaac
     3421 aggatgcatg aagcaacctc tgcagaaact aaacgctcaa agagaaaacg ctgtgaagtc
     3481 tggggagaaa accccaatcc caatgactgg aggagagttg atggttggcc agttgggctg
     3541 aaaaatgttg gcaatacatg ttggtttagt gctgttattc agtctctctt tcaattgcct
     3601 gaatttcgaa gacttgttct cagttatagt ctgccacaaa atgtacttga aaattgtcga
     3661 agtcatacag aaaagagaaa tatcatgttt atgcaagagc ttcagtattt gtttgctcta
     3721 atgatgggat caaatagaaa atttgtagac ccgtctgcag ccctggatct attaaaggga
     3781 gcattccgat catctgagga acagcagcaa gatgtgagtg aattcacaca caagctcctg
     3841 gattggctag aggacgcatt ccagctagct gttaatgtta acagtcccag gaacaaatct
     3901 gaaaatccaa tggtgcagct gttctatggt actttcctga ctgaaggggt tcgtgaagga
     3961 aaaccctttt gtaacaatga gaccttcggc cagtatcctc ttcaggtaaa cggttatcgc
     4021 aacttagacg agtgtttgga aggggccatg gtggagggtg atgttgagct tcttccctcc
     4081 gatcactcgg tgaagtatgg acaagagcgt tggtttacaa agctacctcc agtgttgacc
     4141 tttgaactct caagatttga gtttaatcag tcccttgggc agccagagaa aattcacaat
     4201 aagctggaat ttcctcagat tatttatatg gacaggtaca tgtacaggag caaggagctt
     4261 attcgaaata agagagagtg tattcgaaag ttgaaggagg aaataaaaat tctgcagcaa
     4321 aaattggaaa ggtatgtgaa atatggctca ggcccagctc ggttcccgct cccggacatg
     4381 ctgaaatatg ttattgaatt tgctagtaca aaacctgcct cagaaagctg tccacctgaa
     4441 agtgacacac atatgacatt accactttct tcagtgcact gctcggtttc tgaccagaca
     4501 tccaaggaaa gtacaagtac agaaagctct tctcaggatg ttgaaagtac cttttcttct
     4561 cctgaagatt ctttacccaa gtctaaacca ctgacatctt ctcggtcttc catggaaatg
     4621 ccttcacagc cagctccacg aacagtcaca gatgaggaga taaattttgt taagacctgt
     4681 cttcagagat ggaggagtga gattgaacaa gatatacaag atttaaagac ttgtattgca
     4741 agtactactc agactattga acagatgtac tgcgatcctc tccttcgtca ggtgccttat
     4801 cgcttgcatg cagttcttgt tcatgaagga caagcaaatg ctggacacta ttgggcctat
     4861 atctataatc aaccccgaca gagctggctc aagtacaatg acatctctgt tactgaatct
     4921 tcctgggaag aagttgaaag agattcctat ggaggcctga gaaatgttag tgcttactgt
     4981 ctgatgtaca ttaatgacaa actaccctac ttcaatgcag aggcagcccc aactgaatca
     5041 gatcaaatgt cagaagtgga agccctatct gtggaactca agcattacat tcaggaggat
     5101 aactggcggt ttgagcagga agtagaggag tgggaagaag agcagtcttg caaaatccct
     5161 caaatggagt cctccaccaa ctcctcatca caggactact ctacatcaca agagccttca
     5221 gtagcctctt ctcatggggt tcgctgcttg tcatctgagc atgctgtgat tgtaaaggag
     5281 caaactgccc aggctattgc aaacacagcc cgtgcctatg agaagagcgg tgtagaagcg
     5341 gcactgagtg aggcattcca tgaagaatac tccaggctct atcagcttgc caaagagacc
     5401 cccacctctc acagtgatcc tcgacttcag catgtccttg tctacttttt ccaaaatgaa
     5461 gcacccaaaa gggtagtaga acgaaccctt ctggaacagt ttgcagataa aaatcttagc
     5521 tatgatgaaa gatcaatcag cattatgaag gtggctcaag cgaaactgaa ggaaattggt
     5581 ccagatgaca tgaatatgga agagtacaag aagtggcatg aagattatag tttgttccga
     5641 aaagtgtctg tgtatctcct aacaggccta gaactctatc aaaaaggaaa gtaccaagag
     5701 gcactttcct acctggtata tgcctaccag agcaatgctg ccctgctgat gaaggggccc
     5761 cgccgggggg tcaaagaatc cgtgattgct ttataccgaa gaaaatgcct tctggagctg
     5821 aatgccaaag cagcttctct ttttgaaaca aatgatgatc actccgtaac tgagggcatt
     5881 aatgtgatga atgaactgat catcccctgc attcacctta tcattaataa tgacatttcc
     5941 aaggatgatc tggatgccat tgaggtcatg agaaaccatt ggtgctctta ccttgggcaa
     6001 gatattgcag aaaatctgca gctgtgccta ggggagtttc tacccagact tctagatcct
     6061 tctgcagaaa tcatcgtctt gaaagagcct ccaactattc gacccaattc tccctatgac
     6121 ctatgtagcc gatttgcagc tgtcatggag tcaattcagg gagtttcaac tgtgacagtg
     6181 aaataaaagg gtgggcgcgc cgacccagct ttcttgtaca aagtggttga tatcggtacc
     6241 agtcgactct agaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     6301 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     6361 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     6421 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     6481 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     6541 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     6601 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     6661 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     6721 ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac
     6781 agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg
     6841 tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt
     6901 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6961 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     7021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     7081 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     7141 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     7201 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga
     7261 aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct
     7321 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7381 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7501 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7561 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7621 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata g
//
