LOCUS       CMVTO-KRasG12D          5732 bp ds-DNA     circular     22-JUN-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     polyA_signal    605..735
                     /label="SV40 pA"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     complement(771..791)
                     /label="M13-rev"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_binding    complement(797..819)
                     /label="LacO"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     promoter        complement(823..853)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     rep_origin      1118..1791
                     /label="pUC ori"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     rep_origin      complement(1159..1841)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             complement(1936..2796)
                     /label="Amp (bla)"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     CDS             complement(1939..2598)
                     /label="AmpR"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     promoter        complement(2797..2895)
                     /label="bla promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        3159..3786
                     /label="CMV"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        3238..3541
                     /label="CMV enhancer"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     promoter        3542..3745
                     /label="CMV promoter"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     TATA_signal     3731..3737
                     /label="TATA box"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    3747..3765
                     /label="TetO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    3747..3786
                     /label="TetR binding sites"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    3768..3786
                     /label="TetO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     3910..3917
                     /label="primer3"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    3910..3917
                     /label="Kozak"
                     /ApEinfo_revcolor=#0080ff
                     /ApEinfo_fwdcolor=#0080ff
     CDS             3918..4484
                     /label="Translation 3918-4484"
     misc_feature    3918..4487
                     /label="K-Ras"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    3951..3953
                     /label="G12D"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    4476..4484
                     /label="Cleaved in mature form"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     CDS             4485..4487
                     /label="Translation 4485-4487"
     CDS             4496..4525
                     /label="c-myc epitope"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     misc_feature    4525..4527
                     /label="New Stop"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     CDS             4538..4555
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     misc_feature    4556..4558
                     /label="STOP (missing now)"
                     /ApEinfo_revcolor=#a187be
                     /ApEinfo_fwdcolor=#a187be
     polyA_signal    4577..4791
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     polyA_signal    4824..4847
                     /label="BGH pA"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    5130..5177
                     /label="FRT"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_recomb     5144..5177
                     /label="FRT"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     gene            join(5185..5732,1..473)
                     /label="Hygromycin"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
ORIGIN
        1 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
       61 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
      121 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
      181 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
      241 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
      301 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
      361 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
      421 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
      481 actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
      541 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
      601 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      661 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
      721 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
      781 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      841 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      901 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      961 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1021 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1081 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1141 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1201 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1261 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1321 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1381 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1441 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1501 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1561 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1621 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1681 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1741 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1801 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1861 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1921 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1981 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2041 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2101 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2161 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2221 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2281 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2341 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2401 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2461 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2521 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2581 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2641 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2701 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2761 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2821 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2881 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     2941 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3001 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     3061 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     3121 aggcgttttg cgctgcttcg tacgggccag atatacgcgt tgacattgat tattgactag
     3181 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     3241 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     3301 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     3361 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     3421 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     3481 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     3541 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3601 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     3661 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     3721 gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc ctatcagtga
     3781 tagagatcgt cgacgagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
     3841 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc tagcgtttaa
     3901 acttaagctt cgccaccATG ACAGAGTACA AACTGGTGGT GGTCGGAGCC GACGGCGTGG
     3961 GGAAATCCGC CCTTACCATC CAGCTGATCC AGAATCACTT CGTTGACGAA TATGACCCCA
     4021 CTATAGAGGA CTCTTACCGA AAGCAGGTTG TCATCGACGG CGAAACATGC CTTTTGGACA
     4081 TTCTCGACAC CGCGGGGCAA GAGGAGTATT CTGCAATGAG GGACCAATAC ATGCGCACTG
     4141 GAGAAGGTTT TCTGTGCGTG TTCGCTATTA ATAACACCAA GTCGTTTGAG GATATACACC
     4201 ATTATAGGGA ACAGATTAAA CGCGTGAAGG ATAGCGAAGA TGTACCCATG GTGCTGGTAG
     4261 GTAACAAATG TGATTTACCT AGCAGAACAG TCGACACTAA GCAGGCCCAA GATCTAGCTA
     4321 GGTCCTACGG AATCCCATTC ATCGAGACCA GTGCAAAGAC ACGTCAGAGA GTCGAGGATG
     4381 CCTTTTATAC CCTCGTTAGA GAGATCCGGC AGTACCGGCT GAAAAAGATT TCAAAGGAAG
     4441 AGAAGACGCC GGGGTGCGTG AAAATAAAGA AATGTATTAT CATGtagggc ccGAAcaaaa
     4501 actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcataatc atcattgagt
     4561 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4621 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4681 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4741 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4801 ctctatggag atctatcgat gttctgggga tgcggtgggc tctatggctt ctgaggcgga
     4861 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4921 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4981 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5041 aaatcggggg tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5101 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta
     5161 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     5221 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     5281 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     5341 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     5401 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     5461 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     5521 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     5581 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     5641 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     5701 gtcgcgcagg ctctcgatga gctgatgctt tg
//