LOCUS       Exported                3948 bp ds-DNA     circular SYN 06-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3948)
  AUTHORS   Wythe Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3948
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          185..1840
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          285..326
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          366..371
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          479..1186
                     /organism="mCherry_KRAS4B-WT_GeneBlock"
     source          1467..1472
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1665..1670
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1719..1740
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1735..1740
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    185..284
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     327..358
                     /label=Rox
                     /note="Rox recombination site, Sauer and McDermott 2004"
     regulatory      386..395
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             392..400
                     /codon_start=1
                     /label=Feature 16
                     /translation="MAG"
     CDS             401..421
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             425..445
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             449..469
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             479..1186
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     regulatory      479..482
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(778..799)
                     /label=oJDW 1778
     CDS             1187..1228
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1210..1228)
                     /label=oJDW 1204
     polyA_signal    1270..1405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1275..1466
                     /label=SV40 late polyA
                     /label=SV40 late polyA(1)
     polyA_signal    1284..1405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1473..1664
                     /label=SV40 late polyA
                     /label=SV40 late polyA(1)
     polyA_signal    1473..1603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1482..1603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_recomb     1681..1712
                     /label=Rox
                     /note="Rox recombination site, Sauer and McDermott 2004"
     protein_bind    complement(1741..1840)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1858..1874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1882..1898
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1906..1936)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1951..1972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2260..2848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3026..3835)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacaaata atgattttat tttgactgat agtgacctgt tcgttgcaac acattgatga
      241 gcaatgcttt tttataatgc caactttgta caaaaaagca ggctttaatt aaccgtctta
      301 agcctgtaac aaccggtaca gttcgataac tttaaataat gccaattatt taaagttaca
      361 agcttggtac cctaggttcc cgcgggccac catggccggc ccaaaaaaga agagaaaggt
      421 agatccaaaa aagaagagaa aggtagatcc aaaaaagaag agaaaggtag gatccggcat
      481 ggtgagcaag ggcgaggagg acaacatggc catcatcaag gagttcatgc gcttcaaggt
      541 gcacatggag ggctccgtga acggccacga gttcgagata gaaggagaag gagaaggacg
      601 gccatacgaa ggtacgcaaa cagctaaact caaggtgacc aagggcggcc ccctgccctt
      661 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
      721 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
      781 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tccaggacgg
      841 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
      901 gcagaagaag acgatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
      961 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgccga
     1021 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     1081 catcaagctg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg
     1141 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggta agcctatccc
     1201 taaccctctc ctcggtctcg attctacgta gtaagagcta agcggccgca cttgcaggct
     1261 gcctatatcg ataccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1321 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1381 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1441 tcagggggag gtgtgggagg tttttttcta gacagacatg ataagataca ttgatgagtt
     1501 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1561 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1621 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaagctt ctgcagctag
     1681 taactttaaa taatgccaat tatttaaagt tactcgaggc gatcgccgat atcaacgcgt
     1741 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     1801 acgaacaggt cactatcagt caaaataaaa tcattatttg agcttggtgt aatcatggtc
     1861 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1921 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2461 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2521 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2581 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2641 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2701 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2761 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2821 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2881 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2941 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaagcccaa tctgaataat
     3001 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     3061 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     3121 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     3181 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     3241 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     3301 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     3361 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     3421 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     3481 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt tttccgggga
     3541 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     3601 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     3661 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaagcgat
     3721 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     3781 catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg ctcataacac
     3841 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     3901 cttgtgcaat gtaacatcag agattttgag acacgggcca gagctgca
//
