LOCUS       Exported                6685 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 6685)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6685
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3045..3614
                     /label=NRas_Q61R
     misc_feature    complement(1..188)
                     /label=SV40\polyA\signal
     misc_feature    complement(192..193)
                     /label=MCS 2
     misc_feature    192..193
                     /label=NotI
                     /note="NotI"
     misc_feature    complement(194..241)
                     /label=MCS 2
     misc_feature    complement(264..773)
                     /label=NanoLuc
                     /note="NanoLuc"
     misc_feature    complement(774..788)
                     /label=linker
                     /note="linker"
     misc_feature    complement(788..790)
                     /label=EcoICR I
     misc_feature    complement(795..1076)
                     /label=BRaf RBD aa145-238
                     /note="BRaf RBD aa145-238"
     misc_feature    complement(1078..1085)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    1090..1116
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(1117..1133)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(1136..1181)
                     /label=Minimal CMV Promoter 2
     misc_feature    1159..1181
                     /label=core
                     /note="core"
     promoter        1191
                     /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."
     promoter        1192..1781
                     /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."
     misc_feature    1556..1574
                     /label=HnHc_dR4_2
                     /note="HnHc_dR4_2"
     intron          1896..2028
                     /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        2072..2091
                     /label=T7 promoter
     misc_feature    2106..2996
                     /label=HaloTag
                     /note="Amino Acid sequence is identical to other HT7 
                     versions, but different DNA sequence."
     misc_feature    complement(2146..2166)
                     /label=HTnR
                     /note="HTnR"
     misc_feature    2543..2561
                     /label=HnHc_dR4_3
                     /note="HnHc_dR4_3"
     misc_feature    2997..3035
                     /label=HT7 Linker N terminal
     misc_feature    3009..3029
                     /label=TEV protease recognition sequence
     misc_feature    3036..3043
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    complement(3045..3614)
                     /label=NRas Q61R
                     /note="NRas Q61R"
     misc_feature    3615..3622
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    3654..3669
                     /label=HnHc_dR4_4
                     /note="HnHc_dR4_4"
     misc_feature    3697..3698
                     /label=BlpI
                     /note="BlpI"
     polyA_signal    3718..3939
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(3963..3996)
                     /label=Cre site
                     /note="Cre site"
     promoter        4005..4071
                     /label=EM7 bacterial promoter
     CDS             4085..4879
                     /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."
     misc_feature    4459..4476
                     /label=HnHc_dR4_5
                     /note="HnHc_dR4_5"
     rep_origin      5034..5070
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     misc_feature    5056..5078
                     /label=HnHc_dR4_6
                     /note="HnHc_dR4_6"
     misc_feature    5652..5670
                     /label=HnHc_dR4_7
                     /note="HnHc_dR4_7"
     polyA_signal    complement(5752..5799)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(5823..6221)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(6246..6664)
                     /label=SV40 promoter
                     /note="SV40 promoter"
     misc_feature    6255..6272
                     /label=HnHc_dR4_8
                     /note="HnHc_dR4_8"
     misc_feature    join(6656..6685,1..3)
                     /label=RSGBFB15 (primer)
                     /note="RSGBFB15 (primer)"
ORIGIN
        1 cggcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
       61 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      121 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
      181 gattatgatc cgcggccgca atacgtcgac gttatcagct gacttcgtac gagagcctag
      241 gtgtttctag agtcgcggcc ttacgccaga atgcgttcgc acagccgcca gccggtcact
      301 ccgttgatgg ttactcggaa cagcagggag ccgtcggggt tgatcaggcg ctcgtcgata
      361 attttgttgc cgttccacag ggtccctgtt acagtgatct ttttgccgtc gaacacggcg
      421 atgccttcat acggccgtcc gaaatagtcg atcatgttcg gcgtaacccc gtcgattacc
      481 agtgtgccat agtgcaggat caccttaaag tgatgatcat ccacagggta caccacctta
      541 aaaatttttt cgatctggcc catttggtcg ccgctcagac cttcatacgg gatgatgaca
      601 tggatgtcga tcttcagccc attttcaccg ctcaggacaa tcctttggat cggagttacg
      661 gacaccccga gattctgaaa caaactggac acacctccct gttcaaggac ttggtccagg
      721 ttgtagccgg ctgtctgtcg ccagtcccca acgaaatctt cgagtgtgaa gacgccgctc
      781 gagccgagag aaacaaagtt gtgtgttgta agtggaacat tctccaacac ttccacatgc
      841 aattcttctc cagtaagcca ggaaatatca gtgtcccaac caattggttt cttctctcca
      901 tcctgaattc tgtaaacagc acagcactct gggattagac ctctcatcat cagtgctttc
      961 tttagactgt ctcggactgt aactccacac cttgcaggta ccactgtcct ctgtttgttg
     1021 ggcaggaaga ctctaacgat aggtttttgt ggtgacttgg ggttgctccg tgccatggcg
     1081 atcgctagct gcaaaaagaa caagtagctt gtattctata gtgtcaccta aatgggcgat
     1141 ctgacggttc actaaaccag ctctgcttat ataggcctcc ctggcgcgcc actagttatt
     1201 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     1261 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     1321 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     1381 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc
     1441 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1501 tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     1561 tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     1621 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     1681 caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg
     1741 aggtctatat aagcagagct ggtttagtga accgtcagat cactagaagc tttattgcgg
     1801 tagtttatca cagttaaatt gctaacgcag tcagtgcttc tgacacaaca gtctcgaact
     1861 taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg
     1921 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
     1981 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc
     2041 agttcaatta cagctcttaa ggctagagta ttaatacgac tcactatagg gctagcaaag
     2101 ccaccatggc agaaatcggt actggctttc cattcgaccc ccattatgtg gaagtcctgg
     2161 gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc
     2221 acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc
     2281 atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca gacctgggtt
     2341 atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg ggtctggaag
     2401 aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg gccaagcgca
     2461 atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc ccgacctggg
     2521 acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc
     2581 gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc
     2641 gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc
     2701 gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg
     2761 tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt
     2821 tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc
     2881 tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc
     2941 cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt tccggcgagc
     3001 caaccactga ggatctgtac tttcagagcg ataacgcgat cgccatgact gagtacaaac
     3061 tggtggtggt tggagcaggt ggtgttggga aaagcgcact gacaatccag ctaatccaga
     3121 accactttgt agatgaatat gatcccacca tagaggattc ttacagaaaa caagtggtta
     3181 tagatggtga aacctgtttg ttggacatac tggatacagc tggacgagaa gagtacagtg
     3241 ccatgagaga ccaatacatg aggacaggcg aaggcttcct ctgtgtattt gccatcaata
     3301 atagcaagtc atttgcggat attaacctct acagggagca gattaagcga gtaaaagact
     3361 cggatgatgt acctatggtg ctagtgggaa acaagtgtga tttgccaaca aggacagttg
     3421 atacaaaaca agcccacgaa ctggccaaga gttacgggat tccattcatt gaaacctcag
     3481 ccaagaccag acagggtgtt gaagatgctt tttacacact ggtaagagaa atacgccagt
     3541 accgaatgaa aaaactcaac agcagtgatg atgggactca gggttgtatg ggattgccat
     3601 gtgtggtgat gtaagtttaa acgaattcgg gctcggtacc cggggatcct ctagagtcga
     3661 cctgcaggca tgcaagctga tttggctgct gccaccgctg agcaaggccg cttcgagcag
     3721 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3781 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3841 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     3901 aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcttaatt aagaattcta
     3961 atataacttc gtatagcata cattatacga agttatggat ccactgttga caattaatca
     4021 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca
     4081 gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4141 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4201 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4261 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4321 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4381 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4441 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4501 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4561 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4621 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4681 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4741 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4801 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4861 tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct
     4921 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4981 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5041 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5101 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5161 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5221 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5281 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5341 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5401 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5461 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5521 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5581 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc
     5641 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5701 agattatcaa aaaggatctt cacctagatc cttttatagt ccggaatcgt cacacaaaaa
     5761 accaacacac agatgtaatg aaaataaaga tattttattg cggccatcgt gatggctaga
     5821 gtttagccct cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc
     5881 tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt
     5941 cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa
     6001 gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt
     6061 cccccagtaa aatgatatac attgacacca gtgaagatgc ggccgtcgct agagagagct
     6121 gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt
     6181 tcaatgaggg tggattcttc ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa
     6241 gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     6301 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     6361 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     6421 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     6481 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     6541 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     6601 ccgcctcaga aggtacctaa ccaagttcct ctttcagagg ttatttcagg ccatggtgct
     6661 gcgcaagacg cgttaattaa gacgt
//
