LOCUS       Exported                7430 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Flag-HA-COPS5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7430)
  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 7430)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7430
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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)."
     LTR             359..873
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(949..1548)
                     /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(1032..1052)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1529..1548
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    1560..2112
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1722..1741)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1951..1968
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    2142..2166
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3169..3193)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3209..3235)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3248..3271)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3271..3280
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(3293..3709)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3309..3325)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3341..3363)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3776..4117
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4181..4697
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4897..4916)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5044..5066)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5260..5279)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5379..5401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5439..5457)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5525..5629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5630..6490
                     /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     complement(5848..5867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6661..7249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7150..7169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7403..7420
                     /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 ttggttaaga gatgttaatt tgattaaaca gtttatcctt aataacctga gacatcaatc
     2221 catggatagc ttctatggta gttttacagc tgtctcttgt agctttggca agtttgtctt
     2281 ctgattttcg gtcatgcgtt tctaaaccca acatgaaact ccctcgtccc agctgggctt
     2341 ctgactgctc taacttttca gacaaatcaa agacctgacc agtggtatag tctgcattag
     2401 taagcaagct agaagaactc aacgtattca cccagtattt attccacaac agctcaagca
     2461 atttgcgatc caaagaggat ttgaaatatg agacttctaa ggcataatat tgtttgcagt
     2521 gtacaccaaa atcttctatt ttattaagtg gaatagtctg gtactcagaa ggtccttcat
     2581 caggaggttt gtagcccttt gggtatgtcc taaaggcgcc aagattcact ttccctgcgg
     2641 atattgttct tgttggatca atcaccactg ctacaaatgg ttcctggaac tgctgattga
     2701 gcatctgagt actaacatca atcccagaaa gccagcagcc atagccaggg tggctatgat
     2761 accacccgat tgcattttca aggcggccaa cctgttttgc attttctatg tatgcagcca
     2821 tgtattcata tgcagcagcc tgagcattta ctcgggtttc agtgccctcc acaggcaaag
     2881 caaaactgtc cataatgatc atggtttcac catccacctt tcctagcatc agacccatca
     2941 cttccaagtt gcctcccgat ctggcatgca tcaccatctt cagcagagcc aatgctgaga
     3001 ttttgcagta cttaaagtaa tggtgatcct tagtccaggg cttcgccgcc aggatttctt
     3061 gctgctgttt cttgtcgtat ttgtagattt catcgatact ctgagcttcc tgcatgttgt
     3121 tggccagttc ccaggttttc tgggccatac cgctcccgga cgccgccatg ccaacttttt
     3181 tgtacaaact tgtttgataa cctcgagcag cgtaatctgg aacatcgtat gggtatcctc
     3241 catcgagctt gtcatcgtca tccttgtagt ccatggtggc agatctaatt ccggcgccta
     3301 gagaaggagt gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa
     3361 gggggagaga cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct
     3421 tgacccaggg aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt
     3481 ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc
     3541 ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga
     3601 aggtaaccca acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat
     3661 ctttccagtg acctaaggtc aaacttaagg gagtggtaac agtctggccc taattttcag
     3721 acaaatacag aaacacagtc agacagagac aacacagaac gatgctgcag cgctgcagca
     3781 gacaagacgc gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga ggcgggaact
     3841 gttttaggtt ctcgtctcct accagaacca catatcctga cggggtcgga ttccacatcg
     3901 actcccttcc tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga cgtctcccag
     3961 ggttgcggcc gggtgttcag aactcgtcag ttccaccacg ggtccgccag atacagagct
     4021 agttagctaa ctagtacaga cgcaggcgca aacattagat gccggcacaa acacgcacaa
     4081 agacagagac agacacgaaa cgaccgctgg ccagcttacc tcccggcggg ggggtcggtg
     4141 gtccctgggc aggggtctcc aaatctcggt ggaacctcca aatgaaagac ccccgaggtg
     4201 ggcagtcaat caatctgagg agaccctccc aaggatcagc gagtccacga ttcggatgca
     4261 aacagcaaga ggctttattg ggaatacggg tacccgggcg acgcagtcta tcggaggact
     4321 ggcgcgccga gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga
     4381 acagaagcga gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc
     4441 cttggggcac cctggaaaca tctgatggtt ctctagaaac tgctgagggc gggaccgcat
     4501 ctggggacca tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct
     4561 gtttggccca tattctgctg tctctctgtt cctaaccttg atctgaactt ctctattctc
     4621 agttatgtat tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac
     4681 aggtggggtc tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatct
     4741 atggctcgta ctctataggc ttcagctggt gatattgttg agtcaaaact agagcctgga
     4801 ccactgatat cctgtcttta acaaattgga ctaatcgcct ctacgccgga cgcatcgtgg
     4861 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
     4921 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
     4981 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgc
     5041 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5101 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5161 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     5221 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     5281 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5341 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5401 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5461 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5521 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5581 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5641 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5701 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5761 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5821 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5881 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5941 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6001 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6061 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6121 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6181 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6241 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6301 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6361 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6421 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6481 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6541 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6601 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6661 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6721 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6781 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6841 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6901 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6961 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7021 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7081 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7141 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7201 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7261 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7321 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     7381 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
