LOCUS       73-21_KRAS_G12D_60_AA_l 6404 bp ds-DNA     circular     31-MAY-2024
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    complement(180..227)
                     /label="r"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     modified_base   183..198
                     /label="USER junction"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    184..196
                     /label="Keep unique in all constructs, can be used toexchange antibiotic resistance markers or plasmid origins"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    200..227
                     /label="r"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     rep_origin      complement(215..1041)
                     /label="p15A_origin"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    complement(1016..1042)
                     /label="f"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     modified_base   1043..1057
                     /label="USER junction"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     terminator      1058..1102
                     /label="rrnB1 transcriptional terminator"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    1103..1109
                     /label="nicking endonuclease site that generates overhang"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     modified_base   1110..1124
                     /label="USER junction"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1117..1123
                     /label="Keep unique in all constructs, can be used toexchange f1 origin or selection components"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        1127..1146
                     /label="CGG promoter"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     modified_base   1153..1167
                     /label="USER junction"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    1154..1161
                     /label="SbfI"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    1161..1166
                     /label="BsgI"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     RBS             1167..1182
                     /label="SD8 RBS (from Ringquist and Gold Mol. Micro.1992)"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1183..2457
                     /label="gIII - recoded"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             1183..2457
                     /label="Translation 1183-2457"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTLDRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGGTKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQGALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQGQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKMANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGSNSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVFAFLLYVATFMYVFSTFANILRNKES*"
     misc_feature    2458..2527
                     /label="2xVSV"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    2466..2483
                     /label="VSV term"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    2492..2509
                     /label="VSV term"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    2540..2567
                     /label="kat promoter"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2578..2592
                     /label="SD8 RBS (from Ringquist and Gold Mol. Micro.1992)"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             2593..3156
                     /label="Hs.KRAS4b CDS"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             2593..3156
                     /label="Translation 2593-3156"
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVKDSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREIRKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             2626..2628
                     /label="Hs.KRAS4b CDS"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             2626..2628
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="D"
     misc_feature    3157..3174
                     /label="linker"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             3157..5451
                     /label="Translation 3157-5451"
                     /translation="TSGGSGGSGSSGSGGTSGSGGSGTGTGTGSSGSGGTSGSGGSGSGTGSGSSGSGGTSGSGKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLIESTGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTGITGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANVITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLEFMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYYWLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCFEYAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAKKVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRSVMTLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAVEAMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIKTRVHIMFLGQFEMQPTINTNKDSEIDARKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIHDSFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNLNLRDILESDFAFA*"
     misc_feature    3226..3240
                     /label="overlapping region"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    3337..3364
                     /label="86 Split"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    3337..5451
                     /label="T7 181+"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    4599..4600
                     /label="split 600"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     modified_base   5452..5468
                     /label="USER junction"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    5454..5461
                     /label="Keep unique in all constructs, can be used toexchange antibiotic resistance marker or selectioncomponents"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    5469..5475
                     /label="nicking endonuclease site that generates overhang"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     terminator      complement(5476..5517)
                     /label="termination of bla transcript is slightly weakerthan opposite direction"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     modified_base   5518..5530
                     /label="USER junction"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     CDS             complement(5531..6391)
                     /label="ampR"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     promoter        complement(6392..182)
                     /label="from pUC19"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
ORIGIN
        1 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
       61 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
      121 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
      181 ttaggccaaa taggccgttA GCGTCAGACC CCTTAATAAG ATGATCTTCT TGAGATCGTT
      241 TTGGTCTGCG CGTAATCTCT TGCTCTGAAA ACGAAAAAAC CGCCTTGCAG GGCGGTTTTT
      301 CGAAGGTTCT CTGAGCTACC AACTCTTTGA ACCGAGGTAA CTGGCTTGGA GGAGCGCAGT
      361 CACCAAAACT TGTCCTTTCA GTTTAGCCTT AACCGGCGCA TGACTTCAAG ACTAACTCCT
      421 CTAAATCAAT TACCAGTGGC TGCTGCCAGT GGTGCTTTTG CATGTCTTTC CGGGTTGGAC
      481 TCAAGACGAT AGTTACCGGA TAAGGCGCAG CGGTCGGACT GAACGGGGGG TTCGTGCATA
      541 CAGTCCAGCT TGGAGCGAAC TGCCTACCCG GAACTGAGTG TCAGGCGTGG AATGAGACAA
      601 ACGCGGCCAT AACAGCGGAA TGACACCGGT AAACCGAAAG GCAGGAACAG GAGAGCGCAC
      661 GAGGGAGCCG CCAGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCAC
      721 TGATTTGAGC GTCAGATTTC GTGATGCTTG TCAGGGGGGC GGAGCCTATG GAAAAACGGC
      781 TTTGCCGCGG CCCTCTCACT TCCCTGTTAA GTATCTTCCT GGCATCTTCC AGGAAATCTC
      841 CGCCCCGTTC GTAAGCCATT TCCGCTCGCC GCAGTCGAAC GACCGAGCGT AGCGAGTCAG
      901 TGAGCGAGGA AGCGGAATAT ATCCTGTATC ACATATTCTG CTGACGCACC GGTGCAGCCT
      961 TTTTTCTCCT GCCACATGAA GCACTTCACT GACACCCTCA TCAGTGCCAA CATAGTAAGC
     1021 CAGTATACAC TCCGCTAGCG CTaagaggac atccggtcaa ataaaacgaa aggctcagtc
     1081 gaaagactgg gcctttcgtt ttgctgagga gacttaggga ccctactaat acCGGtcact
     1141 atagggagaa agacctgcag gtgcagtaag gaggaaaaaa aaatgaaaaa attattattc
     1201 gcaattcctt tagttgttcc tttctattct cactccgctg aaactgttga aagttgttta
     1261 gcaaaacccc atacagaaaa ttcatttact aacgtctgga aagacgacaa aactttagat
     1321 cgttacgcta actatgaggg ctgtctgtgg aatgctacag gcgttgtagt ttgtactggt
     1381 gacgaaactc agtgttacgg tacatgggtt cctattgggc ttgctatccc tgaaaatgag
     1441 ggtggtggct ctgagggtgg cggttctgag ggtggcggtt ctgagggtgg cggtactaaa
     1501 cctcctgagt acggtgatac acctattccg ggctatactt atatcaaccc tctcgacggc
     1561 acttatccgc ctggtactga gcaaaacccc gctaatccta atccttctct tgaggagtct
     1621 cagcctctta atactttcat gtttcagaat aataggttcc gaaataggca gggggcatta
     1681 actgtttata cgggcactgt tactcaaggc actgaccccg ttaaaactta ttaccagtac
     1741 actcctgtat catcaaaagc catgtatgac gcttactgga acggtaaatt cagagactgc
     1801 gctttccatt ctggctttaa tgaggatcca ttcgtttgtg aatatcaagg ccaatcgtct
     1861 gacctgcctc aacctcctgt caatgctggc ggcggctctg gtggtggttc tggtggcggc
     1921 tctgagggtg gtggctctga gggtggcggt tctgagggtg gcggctctga gggaggcggt
     1981 tccggtggtg gctctggttc cggtgatttt gattatgaaa agatggcaaa cgctaataag
     2041 ggggctatga ccgaaaatgc cgatgaaaac gcgctacagt ctgacgctaa aggcaaactt
     2101 gattctgtcg ctactgatta cggtgctgct atcgatggtt tcattggtga cgtttccggc
     2161 cttgctaatg gtaatggtgc tactggtgat tttgctggct ctaattccca aatggctcaa
     2221 gtcggtgacg gtgataattc acctttaatg aataatttcc gtcaatattt accttccCTT
     2281 CCACAATCCG TTGAGTGCCG TCCGTTCGTG TTTGGAGCTG GCAAACCCTA TGAGTTCAGC
     2341 ATTGATTGTG ATAAGATCAA TCTGTTTCGT GGGGTATTTG CCTTCTTATT ATATGTCGCA
     2401 ACATTCATGT ATGTGTTTAG CACATTCGCG AACATTCTTC GCAATAAAGA ATCCtaaTAC
     2461 TAGAGTATCT GTTAGTTTTT TTCTACTAGA GTATCTGTTA GTTTTTTTCc atcgtggccg
     2521 gagtacttaa gaagataact aaacaactaa cggacaattc tacctaGGAT CCctcgaaag
     2581 gaggaaaaaa aaatgactga atataaactt gtggtagttg gagctgatgg cgtaggcaag
     2641 agtgccttga cgatacagct aattcagaat cattttgtgg acgaatatga tccaacaata
     2701 gaggattcct acaggaagca agtagtaatt gatggagaaa cctgtctctt ggatattctc
     2761 gacacagcag gtcaagagga gtacagtgca atgagggacc agtacatgag gactggggag
     2821 ggctttcttt gtgtatttgc cataaataat actaaatcat ttgaagatat tcaccattat
     2881 agagaacaaa ttaaaagagt taaggactct gaagatgtac ctatggtcct agtaggaaat
     2941 aaatgtgatt tgccttctag aacagtagac acaaaacagg ctcaggactt agcaagaagt
     3001 tatggaattc cttttattga aacatcagca aagacaagac agggtgttga tgatgccttc
     3061 tatacattag ttcgagaaat tcgaaaacat aaagaaaaga tgagcaaaga tggtaaaaag
     3121 aagaaaaaga agtcaaagac aaagtgtgta attatgACGT CGGGTGGAAG CGGTGGTTCA
     3181 GGGTCCTCAG GTTCGGGTGG AACTAGTGGC TCTGGTGGGT CGGGGACAGG CACAGGGACC
     3241 GGAAGTAGCG GATCGGGTGG CACAAGTGGG TCTGGCGGCT CTGGATCAGG CACCGGCAGT
     3301 GGGTCTTCGG GGAGTGGAGG TACCTCAGGC TCAGGAaaag catttatgca agttgtcgag
     3361 gctgacatgc tctctaaggg tctactcggt ggcgaggcgt ggtcttcgtg gcataaggaa
     3421 gactctattc atgtaggagt acgctgcatc gagatgctca ttgagtcaac cggaatggtt
     3481 agcttacacc gccaaaatgc tggcgtagta ggtcaagact ctgagactat cgaactcgca
     3541 cctgaatacg ctgaggctat cgcaacccgt gcaggtgcgc tggctggcat ctctccgatg
     3601 ttccaacctt gcgtagttcc tcctaagccg tggactggca ttactggtgg tggctattgg
     3661 gctaacggtc gtcgtcctct ggcgctggtg cgtactcaca gtaagaaagc actgatgcgc
     3721 tacgaagacg tttacatgcc tgaggtgtac aaagcgatta acattgcgca aaacaccgca
     3781 tggaaaatca acaagaaagt cctagcggtc gccaacgtaa tcaccaagtg gaagcattgt
     3841 ccggtcgagg acatccctgc gattgagcgt gaagaactcc cgatgaaacc ggaagacatc
     3901 gacatgaatc ctgaggctct caccgcgtgg aaacgtgctg ccgctgctgt gtaccgcaag
     3961 gacaaggctc gcaagtctcg ccgtatcagc cttgagttca tgcttgagca agccaataag
     4021 tttgctaacc ataaggccat ctggttccct tacaacatgg actggcgcgg tcgtgtttac
     4081 gctgtgtcaa tgttcaaccc gcaaggtaac gatatgacca aaggactgct tacgctggcg
     4141 aaaggtaaac caatcggtaa ggaaggttac tactggctga aaatccacgg tgcaaactgt
     4201 gcgggtgtcg ataaggttcc gttccctgag cgcatcaagt tcattgagga aaaccacgag
     4261 aacatcatgg cttgcgctaa gtctccactg gagaacactt ggtgggctga gcaagattct
     4321 ccgttctgct tccttgcgtt ctgctttgag tacgctgggg tacagcacca cggcctgagc
     4381 tataactgct cccttccgct ggcgtttgac gggtcttgct ctggcatcca gcacttctcc
     4441 gcgatgctcc gagatgaggt aggtggtcgc gcggttaact tgcttcctag tgaaaccgtt
     4501 caggacatct acgggattgt tgctaagaaa gtcaacgaga ttctacaagc agacgcaatc
     4561 aatgggaccg ataacgaagt agttaccgtg accgatgaga acactggtga aatctctgag
     4621 aaagtcaagc tgggcactaa ggcactggct ggtcaatggc tggcttacgg tgttactcgc
     4681 agtgtgacta agcgttcagt catgacgctg gcttacgggt ccaaagagtt cggcttccgt
     4741 caacaagtgc tggaagatac cattcagcca gctattgatt ccggcaaggg tctgatgttc
     4801 actcagccga atcaggctgc tggatacatg gctaagctga tttgggaatc tgtgagcgtg
     4861 acggtggtag ctgcggttga agcaatgaac tggcttaagt ctgctgctaa gctgctggct
     4921 gctgaggtca aagataagaa gactggagag attcttcgca agcgttgcgc tgtgcattgg
     4981 gtaactcctg atggtttccc tgtgtggcag gaatacaaga agcctattAA AACGCGCGTG
     5041 CATATTatgt tcctcggtca gttcGAAATG cagcctacca ttaacaccaa caaagatagc
     5101 gagattgatg caCGCaaaca ggagtctggt atcgctccta actttgtaca cagccaagac
     5161 ggtagccacc ttcgtaagac tgtagtgtgg gcacacgaga agtacggaat cgaatctttt
     5221 gcactgattc acgactcctt cggtaccatt ccggctgacg ctgcgaacct gttcaaagca
     5281 gtgcgcgaaa ctatggttga cacatatgag tcttgtgatg tactggctga tttctacgac
     5341 cagttcgctg accagttgca cgagtctcaa ttggacaaaa tgccagcact tccggctaaa
     5401 ggtaacttga acctccgtga catcttagag tcggacttcg cgttcgcgta aacttaatta
     5461 acggcactcc tcagcaaata taatgaccct cttgataacc caagagggca ttttttaatg
     5521 cccatggcgt ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5581 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5641 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5701 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5761 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5821 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5881 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5941 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6001 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6061 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6121 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6181 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6241 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6301 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6361 taagggcgac acggaaatgt tgaatactca tactcttcct tttt
//