LOCUS       Flag-HA-CYLD.        9307 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    Flag-HA-CYLD.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9307)
  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 9307)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9307)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2009 
            Jul 23. 138(2):389-403."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /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)."
     LTR             complement(359..873)
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     CDS             complement(952..1548)
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    complement(1549..2125)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     protein_bind    2142..2166
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2179..5037)
                     /gene="CYLD"
                     /label=CYLD
                     /note="Ubiquitin carboxyl-terminal hydrolase CYLD from Bos 
                     taurus. Accession#: Q1RMU2"
     protein_bind    complement(5046..5070)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5086..5112)
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             complement(5125..5148)
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     regulatory      5148..5157
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5170..5586)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    complement(5653..5994)
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     LTR             complement(6058..6574)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
     primer_bind     complement(6774..6793)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
     primer_bind     complement(6921..6943)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7137..7156)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     7256..7278
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7316..7334)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        7402..7506
                     /label=AmpR promoter
     CDS             7507..8364
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8538..9126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9280..9297
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa
      241 ctatttcagg agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac
      301 ttaatattcg actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat
      361 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      421 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg
      481 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg
      541 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      601 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
      661 tgagggcggg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
      721 cttaccacag atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc
      781 tgaacttctc tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc
      841 tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa
      901 aatcttttat tttatcgata agcttggctg caggtcgaca tcgatgcatc aggcaccggg
      961 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
     1021 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
     1081 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
     1141 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg
     1201 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
     1261 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
     1321 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
     1381 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac
     1441 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
     1501 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg ttgtggccat
     1561 attatcatcg tgtttttcaa aggaaaacca cgtccccgtg gttcgggggg cctagacgtt
     1621 tttttaacct cgactaaaca catgtaaagc atgtgcaccg aggccccaga tcagatccca
     1681 tacaatgggg taccttctgg gcatccttca gccccttgtt gaatacgctt gaggagagcc
     1741 atttgactct ttccacaact atccaactca caacgtggca ctggggttgt gccgcctttg
     1801 caggtgtatc ttatacacgt ggcttttggc cgcagaggca cctgtcgcca ggtggggggt
     1861 tccgctgcct gcaaagggtc gctacagacg ttgtttgtct tcaagaagct tccagaggaa
     1921 ctgcttcctt cacgacattc aacagacctt gcattccttt ggcgagaggg gaaagacccc
     1981 taggaatgct cgtcaagaag acagggccag gtttccgggc cctcacattg ccaaaagacg
     2041 gcaatatggt ggaaaataac atatagacaa acgcacaccg gccttattcc aagcggcttc
     2101 ggccagtaac gttagggggg gggggcggaa ttcgttatcg aaccactttg tacaagaaag
     2161 ttgggtaggc cggccttatt tgtacaaact cattgttgga ctctggtaca tgcacatata
     2221 tgcatcacaa agcagtcttc gtgcacagcc ttggattctc ctggagtcca aggaatgcag
     2281 gtcttccaga gacatcttca agtactctcc tacttctggg catggggtga cttgaggaat
     2341 gttgaagcca ttctgaccac catcccgatc ggccatgctg tcaaagaaga gccaggcaga
     2401 atcgtccttc ccatacttca caaaagcaac atagtggctt gtttctatgc agagaacagc
     2461 aaataactcc atattctggc aagggatgca gccgtgtctc cagtcccagt cgggtaagtc
     2521 tttgggaagt gacactgggt tatatttatg attcagcctc ttcggatgaa ggtggacttg
     2581 agtgttgcag gttttacaaa actgcttgat ttttccagct gagatgtccg gatcgtcgta
     2641 gcattctcta cactcataca ttgcaagccc tccacatatc cggcactgtc tgggagtgtc
     2701 ttcaagtaaa tctgttatat ttaattccag agaaggaaaa atttttttaa atagtttaaa
     2761 gtcttttcca aatcgaggca tctgaataat cagacatgat ggtgcctctg caaatttcag
     2821 gttactgttg ataaaagacc attctaacaa ctgctgaatt gtgggaacgc caactttctc
     2881 atttttttcc ataaaaattt gatagaagta acaatcttgt accttttgac ctgctgatct
     2941 tatttttagc aaaggttcta cccttaaaat atgatgaaac agaatattca agaattcctc
     3001 aggatctttt tcttcagagg taaatcctga tgcagcctcc accttttcaa gtattttcct
     3061 cagtttcata atttttgtgg cacacacata tccatatatt ctcagaggat taacaatttc
     3121 tgtcctcagt agctcttggg tttcactata atattctaca tcgttctttt ctttgggtct
     3181 aagtaacaca gtgtccagaa cagaactaaa agcaaataag cagaataagg ttgagtctaa
     3241 gtaacaagaa ttgtaatgac cctggatgcc tttcttcttc ccaatcatta tctccaagcc
     3301 ttctttttcc atttttggtg gagtattttc ttctactact tcacttaagt agcctccaaa
     3361 tgctaaagag ttacagcgct caatctgatt ggaaaccggc tgcaatgatg caaacctaga
     3421 gtcaggcctg cagctcttca gtttcacaaa cagcgccttc ttcagggcac aggtgaaata
     3481 ccgagtgcct ctgaaggttc catccgtaca gcctgcacac tcatcttcca gttccagtcc
     3541 agcgagcact tcattcagtc ctggtggctg accgatccaa cggattaccc catagaaagg
     3601 agggttctcc ttaacttcag ccaatgagcc cacttctaga ccatgtgagt tcccaggagg
     3661 catggccaag ggtggactct cttggacggg tgcagtgttt agctcttcca ttacagactg
     3721 ggctgacaga gaaagtggac tgtggccaat acttccattg gtattgggca tcttggtgag
     3781 actgaatggt aaagagtgga atctgttctc ggtggtcagt gagttcacag gaggaggctg
     3841 gagtggtggt gaagaacggt caaagtctgt agatatctct gtaagagatt ttgcagggtc
     3901 ttctgcaact tcatcaatgt accatgtatt ttttgatttg gattgtggtt gtgagtcaac
     3961 agaagaccca tttaaggtat aaaataattc agatctgttt ctatttccag ggtctgaggt
     4021 agatcctgta gcctttggtt tattatgact ggatgaacct ttgtccccaa cacctcttga
     4081 cataaaggca agtttgggag gcctcctttc ctgcgtcaca ctctctggga tgatatcatt
     4141 gatgtgcaat agaattgtac tttcaacaca cgcaaaacta caaagctgca ctccatcaaa
     4201 tcttccatcc cagttgccaa tagggttatc catgtccaca ccaacaaaat atcctaagct
     4261 ttcttttcct ggcaaaacat cacagaatat aactgttcca gattctattg tttctccaac
     4321 cttcaaagaa actctggagt ttatttccaa aggaggaagt tcgacctgca ttgtgtcccc
     4381 aggacctgcg taatcacttt ccaatgcagt gtcatcatct tctatgagtt ctagcttgtc
     4441 caatgcaaca aacacgccac aatcttcatc acactgaaaa agctgtttcc cttggtacac
     4501 cccgtcagtg aaaccttgac cacgaccttc ttccagcaat tcaactccaa agaatattcc
     4561 ggagactgtc ctctctgcta acaggggtcc tctgaagcgt acaactccag gaaatttttc
     4621 ttccccagat ctcagctgta ctttcacagg acagcccacg tctatttgga ggcctttact
     4681 tagtctgttt ctgtttttaa acaggctgaa cctctcctca caattggtaa ttgccaaaag
     4741 taactctgtg aacttttcat ttatctctac aacatccttt tcatcaacaa agagaactgc
     4801 atgaggttgc tctagaattt ttaatccaat ctgatttttc ttgccttttg cagaaggaat
     4861 ccttgaatgc cccacagaac gatcttgaat atactgtcct atacttccct tcggtacttt
     4921 aaggagcttt tgtgtttgtt tgtctgtaac gctgcattct tgaagaagca agtaaaaaat
     4981 ccgctcttcc cagtagggtg aagtgacttt ttcttggctc cataagcctg aactcatggt
     5041 gggcgcgcca acttttttgt acaaacttgt ttgataacct cgagcagcgt aatctggaac
     5101 atcgtatggg tatcctccat cgagcttgtc atcgtcatcc ttgtagtcca tggtggcaga
     5161 tctaattccg gcgcctagag aaggagtgag ggctggataa agggaggatc gaggcggggt
     5221 cgaacgagga ggttcaaggg ggagagacgg ggcggatgga ggaagaggag gcggaggctt
     5281 agggtgtaca aagggcttga cccagggagg ggggtcaaaa gccaaggctt cccaggtcac
     5341 gatgtagggg acctggtctg ggtgtccatg cgggccaggt gaaaagacct tgatcttaac
     5401 ctgggtgatg aggtctcggt taaaggtgcc gtctcgcggc catccgacgt taaaggttgg
     5461 ccattctgca gagcagaagg taacccaacg tctcttcttg acatctaccg actggttgtg
     5521 agcgatccgc tcgacatctt tccagtgacc taaggtcaaa cttaagggag tggtaacagt
     5581 ctggccctaa ttttcagaca aatacagaaa cacagtcaga cagagacaac acagaacgat
     5641 gctgcagcgc tgcagcagac aagacgcgcg gcttcggttc caaaccgaaa gcaaaaattc
     5701 agacggaggc gggaactgtt ttaggttctc gtctcctacc agaaccacat atcctgacgg
     5761 ggtcggattc cacatcgact cccttcctca ggtcgggcca caaaaacggc ccccaaagtc
     5821 cctgggacgt ctcccagggt tgcggccggg tgttcagaac tcgtcagttc caccacgggt
     5881 ccgccagata cagagctagt tagctaacta gtacagacgc aggcgcaaac attagatgcc
     5941 ggcacaaaca cgcacaaaga cagagacaga cacgaaacga ccgctggcca gcttacctcc
     6001 cggcgggggg gtcggtggtc cctgggcagg ggtctccaaa tctcggtgga acctccaaat
     6061 gaaagacccc cgaggtgggc agtcaatcaa tctgaggaga ccctcccaag gatcagcgag
     6121 tccacgattc ggatgcaaac agcaagaggc tttattggga atacgggtac ccgggcgacg
     6181 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg
     6241 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
     6301 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
     6361 tgagggcggg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
     6421 cttaccacag atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc
     6481 tgaacttctc tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc
     6541 tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa
     6601 aatcttttat tttatctatg gctcgtactc tataggcttc agctggtgat attgttgagt
     6661 caaaactaga gcctggacca ctgatatcct gtctttaaca aattggacta atcgcctcta
     6721 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     6781 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     6841 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     6901 tgcaccattc cttgcgccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     6961 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     7021 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     7081 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     7141 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     7201 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     7261 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     7321 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     7381 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7441 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7501 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7561 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7621 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7681 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7741 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7801 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7861 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7921 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7981 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     8041 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     8101 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     8161 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     8221 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8281 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8341 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8401 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     9001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     9061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     9121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     9181 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     9241 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     9301 gcggaag
//
