LOCUS       sequence-313756-        7406 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 7406)
  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 7406)
  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-COPS6
FEATURES             Location/Qualifiers
     source          1..7406
                     /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(1520..1544)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1557..2124
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1574..2126
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1718..1735)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1945..1964
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2138..2737
                     /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(2138..2157)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2634..2654
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2813..3327
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3489..3505
                     /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(3513..3519,3520..3537,3538..3543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3558..3579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3696..3713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3867..4455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3947..3966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4626..5417,5418..5486))
                     /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     5249..5268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5487..5591)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5659..5677
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5715..5737)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5837..5856
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6050..6072
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6200..6219
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6419..6935
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6999..7340
                     /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 atggcggcgg cggcggcggc tgcagctacg aacgggaccg gaggaagcag cgggatggag
      601 gtggatgcag cagtagtccc cagcgtgatg gcctgcggag tgactgggag tgtttccgtc
      661 gctctccatc cccttgtcat tctcaacatc tcagaccact ggatccgcat gcgctcccag
      721 gaggggcggc ctgtgcaggt gattggggct ctgattggca agcaggaggg ccgaaatatc
      781 gaggtgatga actcctttga gctgctgtcc cacaccgtgg aagagaagat tatcattgac
      841 aaggaatatt attacaccaa ggaggagcag tttaaacagg tgttcaagga gctggagttt
      901 ctgggttggt ataccacagg ggggccacct gacccctcgg acatccacgt ccataagcag
      961 gtgtgtgaga tcatcgagag ccccctcttt ctgaagttga accctatgac caagcacaca
     1021 gatcttcctg tcagcgtttt tgagtctgtc attgatataa tcaatggaga ggccacaatg
     1081 ctgtttgctg agctgaccta cactctggcc acagaggaag cggaacgcat tggtgtagac
     1141 cacgtagccc gaatgacagc aacaggcagt ggagagaact ccactgtggc tgaacacctg
     1201 atagcacagc acagcgccat caagatgctg cacagccgcg tcaagctcat cttggagtac
     1261 gtcaaggcct ctgaagcggg agaggtcccc tttaatcatg agatcctgcg ggaggcctat
     1321 gctctgtgtc actgtctccc ggtgctcagc acagacaagt tcaagacaga tttttatgat
     1381 caatgcaacg acgtggggct catggcctac ctcggcacca tcaccaaaac gtgcaacacc
     1441 atgaaccagt ttgtgaacaa gttcaatgtc ctctacgacc gacaaggcat cggcaggaga
     1501 atgcgcgggc tctttttctg accaactttc ttgtacaaag tggttcgata acgaattccg
     1561 cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     1621 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     1681 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     1741 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     1801 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     1861 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     1921 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     1981 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     2041 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2101 ttcctttgaa aaacacgatg ataatatggc cacaaccatg accgagtaca agcccacggt
     2161 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     2221 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     2281 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     2341 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     2401 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     2461 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     2521 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     2581 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     2641 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     2701 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatgc atcgatgtcg acctgcagcc
     2761 aagcttatcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     2821 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat
     2881 acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg
     2941 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     3001 gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     3061 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     3121 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     3181 ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc
     3241 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     3301 gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat
     3361 tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc
     3421 acatactcca atactcctga aatagttcat tatggacagc gcagaaagag ctggggagaa
     3481 ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4261 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4321 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4381 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4441 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4501 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4561 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4621 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4681 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4741 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4801 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4861 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4921 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4981 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5041 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5101 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5161 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5221 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5281 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5341 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5401 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5461 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5521 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5581 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5641 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5701 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5761 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5821 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5881 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     5941 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     6001 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     6061 cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     6121 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     6181 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     6241 cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa tttgttaaag
     6301 acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta
     6361 tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     6421 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     6481 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     6541 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     6601 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
     6661 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     6721 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     6781 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
     6841 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
     6901 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
     6961 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
     7021 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
     7081 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
     7141 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
     7201 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     7261 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     7321 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     7381 tgtttctgta tttgtctgaa aattag
//
