LOCUS       sequence-313901-        7690 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 7690)
  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 7690)
  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-STAMBPL1
FEATURES             Location/Qualifiers
     source          1..7690
                     /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"
     protein_bind    complement(1804..1828)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1841..2408
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1858..2410
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2002..2019)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2229..2248
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2422..3021
                     /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(2422..2441)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2918..2938
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3097..3611
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3773..3789
                     /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(3797..3803,3804..3821,3822..3827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3842..3863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3980..3997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4151..4739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4231..4250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4910..5701,5702..5770))
                     /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     5533..5552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5771..5875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5943..5961
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5999..6021)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6121..6140
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6334..6356
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6484..6503
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6703..7219
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7283..7624
                     /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 atgcctgacc atacagatgt ttccctaagc ccagaagagc gagtccgtgc cctaagcaag
      601 cttggttgta atatcaccat cagtgaagac atcactccac gacgttactt taggtctgga
      661 gtagagatgg agaggatggc gtctgtgtat ttggaagaag gaaatttgga aaatgccttt
      721 gttctttata ataaatttat aaccttattt gtagaaaagc ttcctaacca tcgagattac
      781 cagcaatgtg cagtacctga aaagcaggat attatgaaga aactgaagga gattgcattc
      841 ccaaggacag atgaattgaa aaacgacctt ttaaagaaat ataacgtaga ataccaagaa
      901 tatttgcaaa gcaaaaacaa atataaagct gaaattctca aaaaattgga gcatcagaga
      961 ttgatagagg cagaaaggaa gcggattgct cagatgcgcc agcagcagct agaatcggag
     1021 cagtttctgt ttttcgaaga tcaactcaag aagcaagagt tagcccgagg tcaaatgcga
     1081 agtcagcaaa cctcagggct gtcagagcag attgatggga gcgctttgtc ctgcttttcc
     1141 acacaccaga acaattcctt gctgaatgta tttgcagatc aacctaataa aagtgatgca
     1201 accaattatg ctagccactc tcctcctgta aacagggcct taacaccagc tgctactcta
     1261 agtgctgttc agaatttagt ggttgaagga ctgcgatgtg tagttttgcc agaagatctt
     1321 tgccacaaat ttctgcaact ggcagaatct aatacagtga gaggaataga aacctgtgga
     1381 atactctgtg gaaaactgac acataatgaa tttactatta cccatgtaat tgtgccaaag
     1441 cagtctgcgg gaccagacta ttgtgacatg gagaatgtag aggaattatt caatgttcag
     1501 gatcaacatg atctcctcac tctaggatgg atccatacac atcccactca aactgcattt
     1561 ttatccagcg ttgatcttca cactcactgt tcctatcaac tcatgttgcc agaggccatt
     1621 gccattgttt gctcaccaaa gcataaagac actggcatct tcaggctcac caatgctggc
     1681 atgcttgagg tttctgcttg taaaaaaaag ggctttcatc cacacaccaa ggagcccagg
     1741 ctgttcagta tatgcaaaca tgtgttggta aaagacataa aaataattgt gttggattga
     1801 ggtgcccaac tttcttgtac aaagtggttc gataacgaat tccgcccccc ccccctaacg
     1861 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1921 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1981 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     2041 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     2101 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     2161 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     2221 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     2281 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     2341 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     2401 gatgataata tggccacaac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     2461 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     2521 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     2581 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     2641 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     2701 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     2761 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     2821 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     2881 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     2941 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     3001 acccgcaagc ccggtgcctg atgcatcgat gtcgacctgc agccaagctt atcgataaaa
     3061 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     3121 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
     3181 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
     3241 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     3301 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     3361 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     3421 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     3481 gatagactgc gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     3541 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     3601 gggtctttca tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc
     3661 gaatattaag taatcctgac tcaattagcc actgttttga atccacatac tccaatactc
     3721 ctgaaatagt tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc
     3781 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3841 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3901 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3961 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4021 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4081 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4561 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4741 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4801 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4861 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4921 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4981 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5041 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5101 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5161 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5221 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5281 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5341 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5401 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5461 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5521 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5581 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5641 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5701 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5761 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5821 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5881 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5941 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     6001 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     6061 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     6121 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     6181 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     6241 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6301 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6361 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     6421 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     6481 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     6541 cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt
     6601 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
     6661 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     6721 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg
     6781 agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca
     6841 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     6901 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     6961 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     7021 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca
     7081 gtcctccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt
     7141 gcatccgaat cgtggactcg ctgatccttg ggagggtctc ctcagattga ttgactgccc
     7201 acctcggggg tctttcattt ggaggttcca ccgagatttg gagacccctg cccagggacc
     7261 accgaccccc ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt
     7321 tgtgcgtgtt tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt tagctaacta
     7381 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
     7441 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
     7501 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     7561 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     7621 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     7681 tgaaaattag
//
