LOCUS       sequence-357284-        7751 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 7751)
  AUTHORS   Meierhofer D, Wang X, Huang L, Kaiser P
  TITLE     Quantitative analysis of global ubiquitination in HeLa cells by mass
            spectrometry.
  JOURNAL   J Proteome Res. 2008 Oct . 7(10):4566-76.
  PUBMED    18781797
REFERENCE   2  (bases 1 to 7751)
  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: "J Proteome 
            Res. 2008 Oct . 7(10):4566-76."
COMMENT     Sequence Label: pQCXIP HBT-Ubiquitin
FEATURES             Location/Qualifiers
     source          1..7751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             586..761
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    824..1023
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1224..1640
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1570..1592
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1608..1624
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1677..1980
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1981..2184
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2181..2205
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2372..2389
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2642..2662
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2708..2932
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="QIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIFA
                     GKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     misc_feature    3062..3524
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3118..3135)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3345..3364
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3557..4156
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3557..3576)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4053..4073
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4530..4955
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5058..5080)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5216..5236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5221..5550
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5401..5536
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5572..5591
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5672..5689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5843..6431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5923..5942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6602..7393,7394..7462))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7225..7244
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7463..7567)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7635..7653
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcaata aaagagccca
      601 caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt acccgtattc
      661 ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct tgggagggtc
      721 tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc gtccgggatt
      781 tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt
      841 atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg cgtctgtact
      901 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
      961 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     1021 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     1081 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     1141 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     1201 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     1261 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     1321 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     1381 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     1441 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     1501 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     1561 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
     1621 tcactccttc tctaggcgcc ggaattgaag atctggggga tcgatcctct agagtccgtt
     1681 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1741 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1801 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1861 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1921 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1981 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2041 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2101 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2161 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     2221 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
     2281 cggtgcattg gaacggacct gcaggaattg atccgcggcc gcaccggtag gcctcgtacg
     2341 cttaattaac ggatccatta tgaggggttc acatcatcac caccatcatg ctggaaaggc
     2401 cggtgaaggt gaaatccctg cccctcttgc tggtaccgtt tctaagatac tggtaaaaga
     2461 aggtgacact gttaaagctg gtcaaacagt tctggtgctg gaggctatga aaatggagac
     2521 agaaattaac gctcctactg acggaaaagt tgaaaaggtg ttagttaagg aaagagatgc
     2581 tgttcaaggt ggtcaaggtc taatcaagat cggcgactac gatataccca caaccgccag
     2641 cgagaatctg tactttcaat cagagttaaa aaccgccgca ctagctcaac atgacgaggc
     2701 ttctagacag attttcgtca agactttgac cggtaaaacc ataacattgg aagttgaatc
     2761 ttccgatacc atcgacaacg ttaagtcgaa aattcaagac aaggaaggta tccctccaga
     2821 tcaacaaaga ttgatctttg ccggtaagca gctagaagac ggtagaacgc tgtctgatta
     2881 caacattcag aaggagtcca ccttacatct tgtcttaaga ctaagaggtg gttgaggatc
     2941 cggaattccg cccccccccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     3001 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     3061 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     3121 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     3181 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     3241 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     3301 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     3361 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     3421 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     3481 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccac
     3541 aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3601 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3661 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3721 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3781 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3841 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3901 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3961 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     4021 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4081 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4141 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     4201 accccatggc tccgaccgaa gccacccggg gcggccccgc cgaccccgca cccgcccccg
     4261 aggcccaccg actctagtcg agggctgcag cgctgcagag gccgagtgca gaactgctcc
     4321 aaagggacct caaggctttc cgagggacac taggctgact ccatcgagcc agtgtagaga
     4381 taagcttatc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
     4441 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     4501 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     4561 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     4621 ttcagatcaa ggtcaggaac agatggaaca gggtcgaccc tagagaacca tcagatgttt
     4681 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     4741 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     4801 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     4861 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     4921 agtgattgac tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga
     4981 cccctgccca gggaccaccg acccaccacc gggaggtaag ctggctgcct cgcgcgtttc
     5041 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5101 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5161 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtagatccg gctgtggaat
     5221 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5281 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5341 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5401 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5461 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5521 ggcttttttg gaggcctagg cttttgcaaa aagcttactg gcttaactat gcggcatcag
     5581 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     5641 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5701 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5761 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5821 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5881 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5941 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6001 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6061 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6121 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6181 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6241 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     6301 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6361 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6421 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6481 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6541 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6601 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6661 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6721 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6781 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6841 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6901 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6961 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7021 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7081 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7141 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7201 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7261 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7321 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7381 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7441 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7501 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7561 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7621 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttagcttggc
     7681 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat
     7741 taccgccatg t
//
