LOCUS       Exported                5746 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 5746)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5746
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2109..2675
                     /label=KRas_2B_G13D
     misc_feature    complement(1..188)
                     /label=SV40\polyA\signal
     misc_feature    complement(192..254)
                     /label=MCS 2
     misc_feature    192..199
                     /label=NotI
                     /note="NotI"
     misc_feature    197..218
                     /label=HnHc_dR4_1
                     /note="HnHc_dR4_1"
     promoter        255..845
                     /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    620..638
                     /label=HnHc_dR4_2
                     /note="HnHc_dR4_2"
     intron          960..1092
                     /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        1136..1155
                     /label=T7 promoter
     misc_feature    1170..2060
                     /label=HaloTag
                     /note="Amino Acid sequence is identical to other HT7 
                     versions, but different DNA sequence."
     misc_feature    complement(1210..1230)
                     /label=HTnR
                     /note="HTnR"
     misc_feature    1607..1625
                     /label=HnHc_dR4_3
                     /note="HnHc_dR4_3"
     misc_feature    2061..2099
                     /label=HT7 Linker N terminal
     misc_feature    2073..2093
                     /label=TEV protease recognition sequence
     misc_feature    2100..2107
                     /label=SgfI Site
                     /note="SgfI Site"
     misc_feature    2109..2675
                     /label=KRas 2B G13D
                     /note="KRas 2B G13D"
     misc_feature    2676..2683
                     /label=PmeI Site
                     /note="PmeI Site"
     misc_feature    2715..2730
                     /label=HnHc_dR4_4
                     /note="HnHc_dR4_4"
     misc_feature    2758..2759
                     /label=BlpI
                     /note="BlpI"
     polyA_signal    2779..3000
                     /label=Sv40 late polyadenylation signal
     misc_feature    complement(3024..3057)
                     /label=Cre site
                     /note="Cre site"
     promoter        3066..3132
                     /label=EM7 bacterial promoter
     CDS             3146..3940
                     /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    3520..3537
                     /label=HnHc_dR4_5
                     /note="HnHc_dR4_5"
     rep_origin      4095..4131
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     misc_feature    4117..4139
                     /label=HnHc_dR4_6
                     /note="HnHc_dR4_6"
     misc_feature    4713..4731
                     /label=HnHc_dR4_7
                     /note="HnHc_dR4_7"
     polyA_signal    complement(4813..4860)
                     /label=Synthetic PolyAdenylation Signal
     misc_feature    complement(4884..5282)
                     /label=BlastR
                     /note="BlastR"
     misc_feature    complement(5307..5725)
                     /label=SV40 promoter
                     /note="SV40 promoter"
     misc_feature    5316..5333
                     /label=HnHc_dR4_8
                     /note="HnHc_dR4_8"
     misc_feature    join(5717..5746,1..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 gattatggcg cgccactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      841 agatcactag aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      901 cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg
      961 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1021 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     1081 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtattaata
     1141 cgactcacta tagggctagc aaagccacca tggcagaaat cggtactggc tttccattcg
     1201 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     1261 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     1321 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     1381 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     1441 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     1501 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     1561 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     1621 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     1681 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     1741 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     1801 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     1861 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     1921 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     1981 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     2041 cgacgctcga gatttccggc gagccaacca ctgaggatct gtactttcag agcgataacg
     2101 cgatcgccat gactgaatat aaacttgtgg tagttggagc tggtgacgta ggcaagagtg
     2161 ccttgacgat acagctaatt cagaatcatt ttgtggacga atatgatcca acaatagagg
     2221 attcctacag gaagcaagta gtaattgatg gagaaacctg tctcttggat attctcgaca
     2281 cagcaggtca agaggagtac agtgcaatga gggaccagta catgaggact ggggagggct
     2341 ttctttgtgt atttgccata aataatacta aatcatttga agatattcac cattatagag
     2401 aacaaattaa aagagttaag gactctgaag atgtacctat ggtcctagta ggaaataaat
     2461 gtgatttgcc ttctagaaca gtagacacaa aacaggctca ggacttagca agaagttatg
     2521 gaattccttt tattgaaaca tcagcaaaga caagacaggg tgttgatgat gccttctata
     2581 cattagttcg agaaattcga aaacataaag aaaagatgag caaagatggt aaaaagaaga
     2641 aaaagaagtc aaagacaaag tgtgtaatta tgtaagttta aacgaattcg ggctcggtac
     2701 ccggggatcc tctagagtcg acctgcaggc atgcaagctg atttggctgc tgccaccgct
     2761 gagcaaggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2821 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2881 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2941 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     3001 aaatcttaat taagaattct aatataactt cgtatagcat acattatacg aagttatgga
     3061 tccactgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     3121 aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc
     3181 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3241 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3301 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3361 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3421 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3481 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3541 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3601 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3661 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3721 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3781 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3841 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3901 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga ccgaccaagc
     3961 gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4021 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4501 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4681 tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag
     4801 tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt
     4861 gcggccatcg tgatggctag agtttagccc tcccacacat aaccagaggg cagcaattca
     4921 cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga
     4981 tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca
     5041 tttccgatcg cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc
     5101 accacgagtt ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg
     5161 cggccgtcgc tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg
     5221 ttgattgtag ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggcttggcc
     5281 atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag
     5341 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     5401 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     5461 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     5521 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     5581 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     5641 acattccaca gctggttctt tccgcctcag aaggtaccta accaagttcc tctttcagag
     5701 gttatttcag gccatggtgc tgcgcaagac gcgttaatta agacgt
//
