LOCUS       Exported                6786 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4252992
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6786)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6786)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6786
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    319..440
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(330..349)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     384..403
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      555..601
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      631..767
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    812..883
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1109..1697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1189..1208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2197..3406
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3243..3262
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3410..4438
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4465..4513
                     /note="synthetic polyadenylation signal"
     CDS             4779..5639
                     /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(4997..5016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5652..5788
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5940..6011
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      6016..6062
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6080..6128
                     /note="synthetic polyadenylation signal"
     enhancer        6209..6512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6514..6716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6667..6687
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca
       61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat
      121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt
      181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag
      241 ggcaacctta taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc
      301 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
      361 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
      421 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
      481 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
      541 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      601 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
      661 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
      721 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
      781 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
      841 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
      901 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
      961 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     1021 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     1081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1621 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1681 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1801 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1861 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1921 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     1981 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     2041 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     2101 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     2161 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     2221 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     2281 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     2341 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     2401 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     2461 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     2521 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     2581 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     2641 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     2701 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     2761 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     2821 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     2881 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     2941 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     3001 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     3061 taatgcaggg tggggccacc agccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     3121 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     3181 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     3241 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     3301 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     3361 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa
     3421 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     3481 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     3541 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     3601 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     3661 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     3721 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc
     3781 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     3841 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     3901 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     3961 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4021 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4081 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4141 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg
     4201 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     4261 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     4321 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga
     4381 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc
     4441 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     4501 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     4561 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     4621 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     4681 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     4741 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     4801 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4861 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     4921 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     4981 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5041 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     5101 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5161 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     5221 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5281 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     5341 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     5401 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5461 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5521 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5581 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5641 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     5701 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     5761 gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc
     5821 tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg
     5881 gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac
     5941 atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc
     6001 agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     6061 tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg
     6121 attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat
     6181 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     6241 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     6301 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     6361 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     6421 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     6481 acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg
     6541 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     6601 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     6661 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     6721 tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc
     6781 ggccgc
//
