LOCUS       Exported                6653 bp ds-DNA     linear   SYN 30-MAY-2025
DEFINITION  synthetic linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6653)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 30, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6653
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..2169
                     /label=PCR #1 (pF-BiBiTsmC)
                     /note="PCR #1 (pF-BiBiTsmC)"
     misc_feature    1..24
                     /label=BFBRSG15
                     /note="BFBRSG15"
     misc_feature    complement(1..17)
                     /label=BFBRSG16
                     /note="BFBRSG16"
     misc_feature    1..17
                     /label=PCR #2 (pF-4B)
                     /note="PCR #2 (pF-4B)"
     polyA_signal    52..273
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(297..330)
                     /label=Cre site
                     /note="Cre site"
     promoter        339..405
                     /label=EM7 bacterial promoter
     CDS             419..1213
                     /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    1220..1257
                     /label=Untitled
     rep_origin      1368..1404
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(2086..2133)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(2135..2169)
                     /label=BFBRSG14
                     /note="BFBRSG14"
     misc_feature    2145..2998
                     /label=PCR #2 (pF-4B)
                     /note="PCR #2 (pF-4B)"
     misc_feature    2145..2179
                     /label=BFBRSG13
                     /note="BFBRSG13"
     misc_feature    complement(2157..2555)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(2580..2998)
                     /label=SV40 promoter
                     /note="SV40 promoter"
     misc_feature    complement(2973..2998)
                     /label=BFBRSG16
                     /note="BFBRSG16"
     misc_feature    2990..2998
                     /label=BFBRSG15
                     /note="BFBRSG15"
     misc_feature    2999..3212
                     /label=PCR #1 (pF-BiBiTsmC)
                     /note="PCR #1 (pF-BiBiTsmC)"
     misc_feature    2999
                     /label=PCR #1 (pF-BiBiTsmC)
                     /note="PCR #1 (pF-BiBiTsmC)"
     misc_feature    complement(3020..3207)
                     /label=SV40\polyA\signal
     misc_feature    complement(3211..3212)
                     /label=MCS 2
     misc_feature    complement(3213..3260)
                     /label=MCS 2
     misc_feature    complement(3298..3305)
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    complement(3306..3872)
                     /label=KRAS G12V
                     /note="KRAS G12V"
     misc_feature    complement(3874..3881)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    complement(3882..3893)
                     /label=linker
                     /note="linker"
     misc_feature    complement(3894..4406)
                     /label=NanoLuc
                     /note="NanoLuc"
     misc_feature    4420..4446
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(4447..4463)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(4466..4511)
                     /label=Minimal CMV Promoter 2
     misc_feature    4489..4511
                     /label=core
                     /note="core"
     promoter        4521
                     /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        4522..5111
                     /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          5226..5358
                     /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        5402..5421
                     /label=T7 promoter
     misc_feature    5425..5432
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    5434..5685
                     /label=cRaf RBD aa51-134
                     /note="cRaf RBD aa51-134"
     misc_feature    5692..5736
                     /label=HT7 Linker C terminal
     misc_feature    5737..6627
                     /label=HT7 tag C terminal
     misc_feature    6648..6653
                     /label=SbfI
                     /note="SbfI"
ORIGIN
        1 ggcatgcaag ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga
       61 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      121 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      181 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
      241 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatctt aattaagaat tctaatataa
      301 cttcgtatag catacattat acgaagttat ggatccactg ttgacaatta atcatcggca
      361 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
      421 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
      481 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
      541 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
      601 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
      661 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
      721 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
      781 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
      841 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
      901 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
      961 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     1021 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     1081 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     1141 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     1201 cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     1261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1801 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1921 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     1981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2041 tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac
     2101 acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtttag
     2161 ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca
     2221 tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac
     2281 cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg
     2341 ttgccagctg ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca
     2401 gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg
     2461 gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg
     2521 agggtggatt cttcttgaga caaaggcttg gccatggtgg cgctagtgtc agaagaatcg
     2581 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     2641 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     2701 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     2761 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     2821 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     2881 gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct
     2941 cagaaggtac ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgcaa
     3001 gacgcgttaa ttaagacgtc ggcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3061 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3121 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3181 ctacaaatgt ggtatggctg attatgatcc gcggccgcaa tacgtcgacg ttatcagctg
     3241 acttcgtacg agagcctagg tgtttctaga ggatccccgg gtaccgagcc cgaattcgtt
     3301 taaacttaca taattacaca ctttgtcttt gacttctttt tcttcttttt accatctttg
     3361 ctcatctttt ctttatgttt tcgaatttct cgaactaatg tatagaaggc atcatcaaca
     3421 ccctgtcttg tctttgctga tgtttcaata aaaggaattc cataacttct tgctaagtcc
     3481 tgagcctgtt ttgtgtctac tgttctagaa ggcaaatcac atttatttcc tactaggacc
     3541 ataggtacat cttcagagtc cttaactctt ttaatttgtt ctctataatg gtgaatatct
     3601 tcaaatgatt tagtattatt tatggcaaat acacaaagaa agccctcccc agtcctcatg
     3661 tactggtccc tcattgcact gtactcctct tgacctgctg tgtcgagaat atccaagaga
     3721 caggtttctc catcaattac tacttgcttc ctgtaggaat cctctattgt tggatcatat
     3781 tcgtccacaa aatgattctg aattagctgt atcgtcaagg cactcttgcc tacgccaaca
     3841 gctccaacta ccacaagttt atattcagtc atggcgatcg cgccgctcga gcccgccaga
     3901 atgcgttcgc acagccgcca gccggtcact ccgttgatgg ttactcggaa cagcagggag
     3961 ccgtcggggt tgatcaggcg ctcgtcgata attttgttgc cgttccacag ggtccctgtt
     4021 acagtgatct ttttgccgtc gaacacggcg atgccttcat acggccgtcc gaaatagtcg
     4081 atcatgttcg gcgtaacccc gtcgattacc agtgtgccat agtgcaggat caccttaaag
     4141 tgatgatcat ccacagggta caccacctta aaaatttttt cgatctggcc catttggtcg
     4201 ccgctcagac cttcatacgg gatgatgaca tggatgtcga tcttcagccc attttcaccg
     4261 ctcaggacaa tcctttggat cggagttacg gacaccccga gattctgaaa caaactggac
     4321 acacctccct gttcaaggac ttggtccagg ttgtagccgg ctgtctgtcg ccagtcccca
     4381 acgaaatctt cgagtgtgaa gaccatggtg agcgctagct gcaaaaagaa caagtagctt
     4441 gtattctata gtgtcaccta aatgggcgat ctgacggttc actaaaccag ctctgcttat
     4501 ataggcctcc ctggcgcgcc actagttatt aatagtaatc aattacgggg tcattagttc
     4561 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4621 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4681 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4741 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
     4801 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
     4861 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
     4921 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4981 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
     5041 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     5101 accgtcagat cactagaagc tttattgcgg tagtttatca cagttaaatt gctaacgcag
     5161 tcagtgcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg tgaggcactg
     5221 ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
     5281 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
     5341 ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta
     5401 ttaatacgac tcactatagg gctagcgatc gccatgcctt ctaagacaag caacactatc
     5461 cgtgttttct tgccgaacaa gcaaagaaca gtggtcaatg tgcgaaatgg aatgagcttg
     5521 catgactgcc ttatgaaagc actcaaggtg aggggcctgc aaccagagtg ctgtgcagtg
     5581 ttcagacttc tccacgaaca caaaggtaaa aaagcacgct tagattggaa tactgatgct
     5641 gcgtctttga ttggagaaga acttcaagta gatttcctgg atcatgtttc tctcgagcca
     5701 accactgagg atctgtactt tcagagcgat aacgatggat ccgaaatcgg tactggcttt
     5761 ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt
     5821 ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg
     5881 cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga cctgatcggt
     5941 atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt ccgcttcatg
     6001 gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca cgactggggc
     6061 tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg tattgcattt
     6121 atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc ccgcgagacc
     6181 ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca gaacgttttt
     6241 atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat
     6301 taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt cccaaacgag
     6361 ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata catggactgg
     6421 ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt tctgatccca
     6481 ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc
     6541 ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga gatcgcgcgc
     6601 tggctgtcta ctctggagat ttccggttaa tagaattcta gagtcgacct gca
//
