LOCUS       sequence-313842-        7472 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 7472)
  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 7472)
  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-YOD1
FEATURES             Location/Qualifiers
     source          1..7472
                     /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(1586..1610)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1623..2190
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1640..2192
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1784..1801)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2011..2030
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2204..2803
                     /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(2204..2223)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2700..2720
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2879..3393
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3555..3571
                     /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(3579..3585,3586..3603,3604..3609))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3624..3645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3762..3779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3933..4521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4013..4032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4692..5483,5484..5552))
                     /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     5315..5334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5553..5657)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5725..5743
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5781..5803)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5903..5922
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6116..6138
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6266..6285
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6485..7001
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7065..7406
                     /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 atgtttggcc ccgctaaagg tcgccatttt ggagtccacc cggcgcctgg tttccccggc
      601 ggcgtctccc aacaggctgc cgggaccaaa gctggccccg cgggtgcctg gcctgtgggc
      661 agccggaccg acacgatgtg gcggctccgc tgcaaggcca aggacggcac ccatgttttg
      721 caggggctgt ccagccggac ccgggtgcgg gaactccagg gccaaattgc cgccatcacc
      781 gggatcgccc ccggcggtca gcgaatcctc gtcggatacc ctcccgagtg cctggatctc
      841 agcaatgggg ataccattct ggaagacttg cccatccaat ctggtgacat gctgatcatt
      901 gaagaagacc aaaccaggcc cagaagttca cctgcattta ctaaacgtgg tgcttctagt
      961 tacgtcaggg aaactttgcc tgtgcttacc agaaccgtgg tcccagcaga caactcttgc
     1021 ctctttacta gtgtgtacta tgtcgtcgaa ggaggagtct tgaatccagc ttgtgcccct
     1081 gagatgagac gcctcatagc acaaattgta gcaagcgatc cagacttcta tagtgaggca
     1141 atactgggaa aaacaaatca agagtactgt gactggatca aaagggatga cacttgggga
     1201 ggagcaatag agatatcgat tttgtccaag ttttaccaat gtgaaatatg tgtagtggat
     1261 acacagacag taagaattga tcgttttggg gaagatgcag gatataccaa aagggttctg
     1321 cttatttatg atggcatcca ctatgatcca cttcagcgta acttccctga tccagataca
     1381 cctcctctga ccattttctc ctctaatgat gatattgttc ttgtacaagc actggaatta
     1441 gcagatgaag ctagaagaag gagacagttt actgatgtca accgcttcac cctgagatgc
     1501 atggtatgtc agaaaggatt aactggacaa gcagaagcaa gggaacatgc caaggagaca
     1561 ggccatacca actttggaga agtgtgacca actttcttgt acaaagtggt tcgataacga
     1621 attccgcccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     1681 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     1741 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     1801 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     1861 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     1921 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     1981 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2041 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     2101 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2161 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatgaccg agtacaagcc
     2221 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     2281 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     2341 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     2401 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     2461 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     2521 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     2581 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     2641 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     2701 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     2761 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgatgcatcg atgtcgacct
     2821 gcagccaagc ttatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     2881 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     2941 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     3001 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3061 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     3121 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3181 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3241 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     3301 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     3361 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     3421 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     3481 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg
     3541 ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3601 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3661 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3721 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3781 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3841 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3901 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3961 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4021 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4081 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4141 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4201 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4261 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4321 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4381 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4441 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4501 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4561 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4621 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4681 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4741 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4801 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4861 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4921 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4981 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5041 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5101 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5161 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5221 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5281 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5341 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5401 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5461 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5521 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5581 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5641 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5701 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     5761 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5821 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5881 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     5941 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     6001 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6061 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6121 tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca
     6181 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6241 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6301 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg
     6361 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     6421 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
     6481 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     6541 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     6601 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     6661 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     6721 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     6781 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     6841 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
     6901 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
     6961 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
     7021 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
     7081 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
     7141 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
     7201 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
     7261 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
     7321 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
     7381 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
     7441 tgactgtgtt tctgtatttg tctgaaaatt ag
//
