LOCUS       sequence-159428-        8785 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 8785)
  AUTHORS   Cummins JM, Vogelstein B
  TITLE     HAUSP is required for p53 destabilization.
  JOURNAL   Cell Cycle. 2004 Jun . 3(6):689-92.
  PUBMED    15118411
REFERENCE   2  (bases 1 to 8785)
  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: "Cell Cycle.
            2004 Jun . 3(6):689-92."
COMMENT     Sequence Label: pCI-neo Flag HAUSP
FEATURES             Location/Qualifiers
     source          1..8785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1094..1117
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4489..4610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4500..4519)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4554..4573
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4796..5251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4883..4902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5093..5114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5368..5725
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5368..5559
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5391..5411)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5576..5711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5776..6570
                     /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(5830..5849)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6440..6459
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6634..6682
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6723..6742)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6842..6864
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6902..6920)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6988..7092
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7093..7161,7162..7953)
                     /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     complement(7311..7330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8124..8712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8613..8632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc atggactaca aagacgatga cgacaagatg aaccaccagc agcagcagca
     1141 gcagcagaaa gcgggcgagc agcagttgag cgagcccgag gacatggaga tggaagcggg
     1201 agatacagat gacccaccaa gaattactca gaaccctgtg atcaatggga atgtggccct
     1261 gagtgatgga cacaacaccg cggaggagga catggaggat gacaccagtt ggcgctccga
     1321 ggcaaccttt cagttcactg tggagcgctt cagcagactg agtgagtcgg tccttagccc
     1381 tccgtgtttt gtgcgaaatc tgccatggaa gattatggtg atgccacgct tttatccaga
     1441 cagaccacac caaaaaagcg taggattctt tctccagtgc aatgctgaat ctgattccac
     1501 gtcatggtct tgccatgcac aagcagtgct gaagataata aattacagag atgatgaaaa
     1561 gtcgttcagt cgtcgtatta gtcatttgtt cttccataaa gaaaatgatt ggggattttc
     1621 caattttatg gcctggagtg aagtgaccga tcctgagaaa ggatttatag atgatgacaa
     1681 agttaccttt gaagtctttg tacaggcgga tgctccccat ggagttgcgt gggattcaaa
     1741 gaagcacaca ggctacgtcg gcttaaagaa tcagggagcg acttgttaca tgaacagcct
     1801 gctacagacg ttatttttca cgaatcagct acgaaaggct gtgtacatga tgccaaccga
     1861 gggggatgat tcgtctaaaa gcgtcccttt agcattacaa agagtgttct atgaattaca
     1921 gcatagtgat aaacctgtag gaacaaaaaa gttaacaaag tcatttgggt gggaaacttt
     1981 agatagcttc atgcaacatg atgttcagga gctttgtcga gtgttgctcg ataatgtgga
     2041 aaataagatg aaaggcacct gtgtagaggg caccataccc aaattattcc gcggcaaaat
     2101 ggtgtcctat atccagtgta aagaagtaga ctatcggtct gatagaagag aagattatta
     2161 tgatatccag ctaagtatca aaggaaagaa aaatatattt gaatcatttg tggattatgt
     2221 ggcagtagaa cagctcgatg gggacaataa atacgacgct ggggaacatg gcttacagga
     2281 agcagagaaa ggtgtgaaat tcctaacatt gccaccagtg ttacacctac aactgatgag
     2341 atttatgtat gaccctcaga cggaccaaaa tatcaagatc aatgataggt ttgaattccc
     2401 agagcagtta ccacttgatg aatttttgca aaaaacagat cctaaggacc ctgcaaatta
     2461 tattcttcat gcagtcctgg ttcatagtgg agataatcat ggtggacatt atgtggttta
     2521 tctaaacccc aaaggggatg gcaaatggtg taaatttgat gacgacgtgg tgtcaaggtg
     2581 tactaaagag gaagcaattg agcacaatta tgggggtcac gatgacgacc tgtctgttcg
     2641 acactgcact aatgcttaca tgttagtcta catcagggaa tcaaaactga gtgaagtttt
     2701 acaggcggtc accgaccatg atattcctca gcagttggtg gagcgattac aagaagagaa
     2761 aaggatcgag gctcagaagc ggaaggagcg gcaggaagcc catctctata tgcaagtgca
     2821 gatagtcgca gaggaccagt tttgtggcca ccaagggaat gacatgtacg atgaagaaaa
     2881 agtgaaatac actgtgttca aagtattgaa gaactcctcg cttgctgagt ttgttcagag
     2941 cctctctcag accatgggat ttccacaaga tcaaattcga ttgtggccca tgcaagcaag
     3001 gagtaatgga acaaaacgac cagcaatgtt agataatgaa gccgacggca ataaaacaat
     3061 gattgagctc agtgataatg aaaacccttg gacaatattc ctggaaacag ttgatcccga
     3121 gctggctgct agtggagcga ccttacccaa gtttgataaa gatcatgatg taatgttatt
     3181 tttgaagatg tatgatccca aaacgcggag cttgaattac tgtgggcata tctacacacc
     3241 aatatcctgt aaaatacgtg acttgctccc agttatgtgt gacagagcag gatttattca
     3301 agatactagc cttatcctct atgaggaagt taaaccgaat ttaacagaga gaattcagga
     3361 ctatgacgtg tctcttgata aagcccttga tgaactaatg gatggtgaca tcatagtatt
     3421 tcagaaggat gaccctgaaa atgataacag tgaattaccc accgcaaagg agtatttccg
     3481 agatctctac caccgcgttg atgtcatttt ctgtgataaa acaatcccta atgatcctgg
     3541 atttgtggtt acgttatcaa atagaatgaa ttattttcag gttgcaaaga cagttgcaca
     3601 gaggctcaac acagatccaa tgttgctgca gtttttcaag tctcaaggtt atagggatgg
     3661 cccaggtaat cctcttagac ataattatga aggtacttta agagatcttc tacagttctt
     3721 caagcctaga caacctaaga aactttacta tcagcagctt aagatgaaaa tcacagactt
     3781 tgagaacagg cgaagtttta aatgtatatg gttaaacagc caatttaggg aagaggaaat
     3841 aacactatat ccagacaagc atgggtgtgt ccgggacctg ttagaagaat gtaaaaaggc
     3901 cgtggagctt ggggagaaag catcagggaa acttaggctg ctagaaattg taagctacaa
     3961 aatcattggt gttcatcaag aagatgaact attagaatgt ttatctcctg caacgagccg
     4021 gacgtttcga atagaggaaa tccctttgga ccaggtggac atagacaaag agaatgagat
     4081 gcttgtcaca gtggcgcatt tccacaaaga ggtcttcgga acgttcggaa tcccgttttt
     4141 gctgaggata caccagggcg agcattttcg agaagtgatg aagcgaatcc agagcctgct
     4201 ggacatccag gagaaggagt ttgagaagtt taaatttgca attgtaatga tgggccgaca
     4261 ccagtacata aatgaagacg agtatgaagt aaatttgaaa gactttgagc cacagcccgg
     4321 taatatgtct catcctcggc cttggctagg gctcgaccac ttcaacaaag ccccaaagag
     4381 gagtcgctac acttaccttg aaaaggccat taaaatccat aactgatcta gagtcgaccc
     4441 gggcggccgc ttccctttag tgagggttaa tgcttcgagc agacatgata agatacattg
     4501 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4561 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4621 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     4681 aaaacctcta caaatgtggt aaaatccgat aaggatcgat ccgggctggc gtaatagcga
     4741 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     4801 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4861 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4921 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4981 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5041 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5101 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5161 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5221 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     5281 tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc
     5341 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     5401 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5461 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5521 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5581 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5641 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5701 ttttggaggc ctaggctttt gcaaaaagct tgattcttct gacacaacag tctcgaactt
     5761 aaggctagag ccaccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5821 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5881 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5941 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6001 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     6061 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     6121 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6181 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6241 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6301 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6361 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6421 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6481 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6541 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6601 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     6661 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgcgtatg gtgcactctc
     6721 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6781 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6841 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     6901 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6961 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     7021 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     7081 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     7141 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     7201 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7261 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7321 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     7381 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7441 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7501 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7561 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7621 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7681 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7741 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7801 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7861 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7921 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7981 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     8041 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     8101 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     8161 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     8221 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     8281 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     8341 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     8401 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     8461 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8521 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8581 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8641 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8701 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8761 tttgctcaca tggctcgaca gatct
//
