LOCUS       Exported                4898 bp ds-DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG195 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTsmN w/ BFBRSG15/BFBRSG14 (PCR #1; 3.9 kbp);

            PCR amplify pF-4B w/ BFBRSG13/BFBRSG16 (PCR #2; 871 bp); 

            2X component Gibson Assembly;

            electroporation of BR610.
FEATURES             Location/Qualifiers
     source          1..4898
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1279..1845
                     /label=NRas_WT_NM_002524
     misc_feature    complement(22..209)
                     /label=SV40\polyA\signal
     misc_feature    complement(213..275)
                     /label=MCS 2
     promoter        276..866
                     /label=CMV promoter
                     /note="Has EcoICRI site removed. Named changed from ""CMV 
                     Promoter"" to ""CMV immediate-early enhancer/promoter"" 
                     3/21/2011 and J Hartnet's request to fit Sci Com's naming."
     intron          981..1113
                     /label=chimeric intron
                     /note="Note that this is the actual sequence spliced out. 
                     The 5' donor site includes a 5' GA and the 3' acceptor site
                     adds a 3'G."
     promoter        1157..1176
                     /label=T7 promoter
     CDS             1189..1224
                     /codon_start=1
                     /label=SmBiT
                     /note="SmBiT"
                     /translation="MVTGYRLFEEIL"
     CDS             1225..1269
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     misc_feature    1270..1277
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1279..1845
                     /label=NRAS (Q61R)
                     /note="NRAS (Q61R)"
     misc_feature    1459..1461
                     /label=Q61R
                     /note="Q61R"
     misc_feature    1849..1856
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    1952..2173
                     /label=Sv40 late polyadenylation signal
     promoter        2239..2305
                     /label=EM7 bacterial promoter
     CDS             2319..3113
                     /label=KanR
                     /note="Name changed frm ""Neomycin phosphotransferase"" to 
                     ""Neomycin phosphotransferase coding region"" on 3/21/2011 
                     at Jim Hartnet's request to fit Sci Com's naming."
     rep_origin      3268..3304
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(3986..4033)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4057..4455)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(4480..4898)
                     /label=SV40 promoter
                     /note="SV40 promoter"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gactgagtac aaactggtgg tggttggagc aggtggtgtt
     1321 gggaaaagcg cactgacaat ccagctaatc cagaaccact ttgtagatga atatgatccc
     1381 accatagagg attcttacag aaaacaagtg gttatagatg gtgaaacctg tttgttggac
     1441 atactggata cagctggaag agaagagtac agtgccatga gagaccaata catgaggaca
     1501 ggcgaaggct tcctctgtgt atttgccatc aataatagca agtcatttgc ggatattaac
     1561 ctctacaggg agcagattaa gcgagtaaaa gactcggatg atgtacctat ggtgctagtg
     1621 ggaaacaagt gtgatttgcc aacaaggaca gttgatacaa aacaagccca cgaactggcc
     1681 aagagttacg ggattccatt cattgaaacc tcagccaaga ccagacaggg tgttgaagat
     1741 gctttttaca cactggtaag agaaatacgc cagtaccgaa tgaaaaaact caacagcagt
     1801 gatgatggga ctcagggttg tatgggattg ccatgtgtgg tgatgtaagt ttaaacgaat
     1861 tcgggctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag ctgatttggc
     1921 tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat tgatgagttt
     1981 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2041 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2101 cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc
     2161 tacaaatgtg gtaaaatctt aattaagaat tctaatataa cttcgtatag catacattat
     2221 acgaagttat ggatccactg ttgacaatta atcatcggca tagtatatcg gcatagtata
     2281 atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc
     2341 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2401 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2461 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     2521 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2581 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2641 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2701 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2761 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2821 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     2881 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2941 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3001 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3061 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaaa
     3121 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     3181 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3241 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3301 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3361 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3421 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3481 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3541 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3601 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3661 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3721 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3781 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3841 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3901 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3961 gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt aatgaaaata
     4021 aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca cataaccaga
     4081 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     4141 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     4201 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     4261 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     4321 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     4381 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     4441 caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag
     4501 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     4561 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     4621 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     4681 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     4741 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     4801 ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac ctaaccaagt
     4861 tcctctttca gaggttattt caggccatgg tgctgcgc
//
