LOCUS       sequence-313899-        7400 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 7400)
  AUTHORS   Sowa ME, Bennett EJ, Gygi SP, Harper JW
  TITLE     Defining the human deubiquitinating enzyme interaction landscape.
  JOURNAL   Cell. 2009 Jul 23. 138(2):389-403.
  PUBMED    19615732
REFERENCE   2  (bases 1 to 7400)
  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. 2009 
            Jul 23. 138(2):389-403."
COMMENT     Sequence Label: Flag-HA-PSMD7
FEATURES             Location/Qualifiers
     source          1..7400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..417
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     347..369
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     385..401
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      430..439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             439..462
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             475..501
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    517..541
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             1489..1500
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(1514..1538)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1551..2118
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1568..2120
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1712..1729)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1939..1958
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2132..2731
                     /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(2132..2151)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2628..2648
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2807..3321
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3483..3499
                     /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(3507..3513,3514..3531,3532..3537))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3552..3573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3690..3707)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3861..4449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3941..3960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4620..5411,5412..5480))
                     /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     5243..5262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5481..5585)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5653..5671
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5709..5731)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5831..5850
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6044..6066
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6194..6213
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6413..6929
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6993..7334
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
ORIGIN
        1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
       61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
      421 attagatctg ccaccatgga ctacaaggat gacgatgaca agctcgatgg aggataccca
      481 tacgatgttc cagattacgc tgctcgaggt tatcaaacaa gtttgtacaa aaaagttggc
      541 atgccggagc tggcagtgca gaaggtggtg gtccaccccc tggtgctgct cagtgtggtg
      601 gatcatttca accgaatcgg caaggttgga aaccagaagc gtgttgttgg tgtgcttttg
      661 gggtcatggc aaaagaaagt acttgatgta tcgaacagtt ttgcagttcc ttttgatgaa
      721 gatgacaaag acgattctgt atggttttta gaccatgatt atttggaaaa catgtatgga
      781 atgtttaaga aagtcaatgc cagggaaaga atagttggct ggtaccacac aggccctaaa
      841 ctacacaaga atgacattgc catcaacgaa ctcatgaaaa gatactgtcc taattccgta
      901 ttggtcatca ttgatgtgaa gccgaaggac ctagggctgc ctacagaagc gtacatttca
      961 gtggaagaag tccatgatga tggaactcca acctcgaaaa catttgaaca cgtgaccagt
     1021 gaaattggag cagaggaagc tgaggaagtt ggagttgaac acttgttacg agatatcaaa
     1081 gacacgacgg tgggcactct gtcccagcgg atcacaaacc aggtccatgg tttgaaggga
     1141 ctgaactcca agcttctgga tatcaggagc tacctggaaa aagtcgccac aggcaagctg
     1201 cccatcaacc accagatcat ctaccagctg caggacgtct tcaacctgct gccagatgtc
     1261 agcctgcagg agttcgtcaa ggccttttac ctgaagacca atgaccagat ggtggtagtg
     1321 tacttggcct cgctgatccg ttccgtggtc gccctgcaca acctcatcaa caacaagatt
     1381 gccaaccggg atgcagagaa gaaagaaggg caggagaaag aagagagcaa aaaggatagg
     1441 aaagaggaca aggagaaaga taaagataag gaaaagagtg atgtaaagaa agaggagaaa
     1501 aaggagaaaa agtaaccaac tttcttgtac aaagtggttc gataacgaat tccgcccccc
     1561 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     1621 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     1681 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     1741 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     1801 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     1861 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     1921 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     1981 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     2041 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     2101 tgaaaaacac gatgataata tggccacaac catgaccgag tacaagccca cggtgcgcct
     2161 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     2221 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     2281 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     2341 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     2401 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     2461 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     2521 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     2581 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     2641 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     2701 ctggtgcatg acccgcaagc ccggtgcctg atgcatcgat gtcgacctgc agccaagctt
     2761 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     2821 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa
     2881 ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa
     2941 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     3001 agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     3061 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     3121 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg
     3181 ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     3241 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     3301 ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca acattctgag
     3361 ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga atccacatac
     3421 tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg agaattgtga
     3481 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3541 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3601 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3661 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3721 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3781 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3841 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3901 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3961 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4021 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4081 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4141 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4201 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4261 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4321 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4381 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4441 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4501 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4561 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4621 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4681 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4741 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4801 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4861 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4921 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4981 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5041 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5101 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5161 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5221 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5281 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5341 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5401 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5461 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5521 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5581 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     5641 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     5701 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5761 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     5821 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     5881 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     5941 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     6001 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     6061 aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     6121 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     6181 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6241 tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga
     6301 tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt
     6361 acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     6421 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     6481 tacataactg agaatagaga agttcagatc aaggttagga acagagagac agcagaatat
     6541 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     6601 ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg
     6661 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     6721 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     6781 cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
     6841 cttgctgttt gcatccgaat cgtggactcg ctgatccttg ggagggtctc ctcagattga
     6901 ttgactgccc acctcggggg tctttcattt ggaggttcca ccgagatttg gagacccctg
     6961 cccagggacc accgaccccc ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg
     7021 tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt
     7081 tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg
     7141 gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag
     7201 gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga
     7261 acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg
     7321 cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc
     7381 tgtatttgtc tgaaaattag
//
