pRK5-HA-Ubiquitin-K6

LOCUS       pRK5-HA-Ubiquitin-K6               4996 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pRK5-HA-Ubiquitin-K6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4996)
  AUTHORS   Livingston CM, Ifrim MF, Cowan AE, Weller SK
  TITLE     Virus-Induced Chaperone-Enriched (VICE) domains function as nuclear 
            protein quality control centers during HSV-1 infection.
  JOURNAL   PLoS Pathog. 2009 Oct . 5(10):e1000619.
  PUBMED    19816571
REFERENCE   2  (bases 1 to 4996)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..4996
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    98..119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        134..164
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    172..188
                     /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)."
     primer_bind     177..199
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     196..212
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     196..212
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        241..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     821..845
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1055..1073
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1055..1072
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1190..1199
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1202..1228
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1259..1486
                     /codon_start=1
                     /product="human ubiquitin variant containing Lys-6, with 
                     all other lysines mutated to arginine"
                     /label=ubiquitin-K6
                     /translation="MQIFVKTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQRESTLHLVLRLRGG"
     regulatory      1523..1532
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    1536..1670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1573..1592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1627..1646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1739..2096
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1762..1782)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1947..2082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2009..2028
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2116..2133)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2116..2132)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2125..2147)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2345..2800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2432..2451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2642..2663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2817..2836)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2936..2958
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2996..3014)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3082..3186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3187..4047
                     /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(3405..3424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4218..4806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4707..4726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4960..4977
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
       61 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      121 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      181 gataacaatt tcacacagga aacagctatg acatgattac gaattaattc gagctcgccc
      241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      841 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      901 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      961 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
     1021 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
     1141 cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg ccaccatggg
     1201 ctacccctat gatgtgcctg actacgcaga tctcaatggt ggtggtggtg ggtcgaccat
     1261 gcagatcttc gtcaaaacgt taaccggtag aaccataact ctagaagttg aaccatccga
     1321 taccatcgaa aacgttagag ctagaattca agacagagaa ggcattccac ctgatcaaca
     1381 aagattgatc tttgccggta gacagctcga ggacggtaga acgctgtctg attacaacat
     1441 tcagagagag tcgaccttac atcttgtctt aagactaaga ggtggttgag cggccgctaa
     1501 gtaagtaagg atccccagct tggccgccat ggcccaactt gtttattgca gcttataatg
     1561 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1621 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattaat
     1681 tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct
     1741 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     1861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1921 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1981 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2041 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2101 tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2161 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2221 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     2281 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     2341 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2401 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2461 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2521 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2581 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2641 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttaat
     2701 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2761 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     2821 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2881 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2941 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     3001 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3061 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3301 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3361 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3421 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3481 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3541 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3601 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3661 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3721 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3781 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3841 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3901 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3961 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4021 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4081 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4141 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4921 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4981 gcggaagagc gcccaa
//