LOCUS       pRK5-HA-Ubiquitin-K29                4996 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pRK5-HA-Ubiquitin-K29
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
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..4996
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        264..294
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /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     307..329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        371..750
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        751..954
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     905..925
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     951..975
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1185..1203
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1185..1202
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1216..1235
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1320..1329
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1332..1358
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1389..1616
                     /codon_start=1
                     /product="human ubiquitin variant containing Lys-29, with 
                     all other lysines mutated to arginine"
                     /label=ubiquitin-K29
                     /translation="MQIFVRTLTGRTITLEVEPSDTIENVRAKIQDREGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQRESTLHLVLRLRGG"
     regulatory      1653..1662
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    1666..1800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1703..1722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1757..1776
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1869..2226
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1892..1912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2077..2212
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2139..2158
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2246..2263)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2246..2262)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2255..2277)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2475..2930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2562..2581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2772..2793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2947..2966)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3066..3088
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3126..3144)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3212..3316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3317..4177
                     /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(3535..3554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4348..4936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4837..4856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattaattc
      361 gagctcgccc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      421 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      481 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      541 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      601 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      661 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      721 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      781 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      841 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      901 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      961 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     1021 cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc
     1081 aagagtgacg taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa
     1141 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     1201 agaataacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc
     1261 tcggttctat cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg
     1321 ccaccatggg ctacccctat gatgtgcctg actacgcaga tctcaatggt ggtggtggtg
     1381 ggtcgaccat gcagatcttc gtcagaacgt taaccggtag aaccataact ctagaagttg
     1441 aaccatccga taccatcgaa aacgttagag ctaaaattca agacagagaa ggcattccac
     1501 ctgatcaaca aagattgatc tttgccggta gacagctcga ggacggtaga acgctgtctg
     1561 attacaacat tcagagagag tcgaccttac atcttgtctt aagactaaga ggtggttgag
     1621 cggccgctaa gtaagtaagg atccccagct tggccgccat ggcccaactt gtttattgca
     1681 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1741 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     1801 gggaattaat tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     1861 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     1921 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1981 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2041 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2101 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2161 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2221 tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     2281 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2341 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2401 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2461 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2521 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2581 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2641 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2701 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2761 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     2821 ttcttttaat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2881 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2941 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3001 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3061 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3121 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3181 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3241 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3301 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3361 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3421 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3481 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3541 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3601 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3661 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3721 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3781 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3841 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3901 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3961 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4021 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4081 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4141 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4201 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4261 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4321 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4381 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4441 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4501 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4561 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4621 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4681 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4741 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4801 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4861 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4921 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4981 gccttttgct cacatg
//