LOCUS       sequence-368438-        4803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eGFP-KRAS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4803)
  AUTHORS   Shi Z, Graber ZT, Baumgart T, Stone HA, Cohen AE
  TITLE     Cell Membranes Resist Flow.
  JOURNAL   Cell. 2018 Dec 13;175(7):1769-1779.e13. doi: 
            10.1016/j.cell.2018.09.054. Epub 2018 Nov 1.
  PUBMED    30392960
REFERENCE   2  (bases 1 to 4803)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2018.09.054"; journalName: "Cell"; date: 
            "2018-12-13- 13"; volume: "175"; issue: "7"; pages: "1769-1779.e13"
COMMENT     Sequence Label: ZS006:eGFP-KRAS
FEATURES             Location/Qualifiers
     source          1..4803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             168..962
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             168..962
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(222..241)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     832..851
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1150..1169)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1194..1241
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1570..2158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2059..2078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2333..2636
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2637..2840
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2791..2811
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2879..2888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2885..2887,2888..2890,2891..3601)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2885..2887,2888..2890,2891..3601)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2930..2951)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3191..3210)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3538..3559
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3863..3984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3900..3919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3954..3973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3991..4446)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4128..4149)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4340..4359
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4473..4577
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4579..4803,1..133)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4579..4770
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4602..4622)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(4787..4803,1..119)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
       61 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
      121 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
      181 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
      241 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
      301 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
      361 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
      421 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
      481 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
      541 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
      601 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
      661 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
      721 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
      781 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
      841 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
      901 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
      961 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     1021 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     1081 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     1141 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     1201 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     1261 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     1321 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     1381 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     1441 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1501 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1561 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1621 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1681 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1741 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1801 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1861 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1921 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1981 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2041 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2101 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2161 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2221 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     2281 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     2341 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     2401 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     2461 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     2521 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     2581 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     2641 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     2701 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     2761 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     2821 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc
     2881 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2941 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     3001 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     3061 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     3121 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3181 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3241 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3301 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3361 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3421 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3481 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3541 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3601 gtccggactc agatctcgag ctcaagcttg gggtaaaaag aagaaaaaga agtcaaagac
     3661 aaagtgtgta attatgtaat gagaattcga tatcaagctt atcgatactg cagtcgacgg
     3721 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     3781 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3841 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3901 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3961 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     4021 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4081 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4141 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4201 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4261 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4321 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4381 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4441 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4501 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4561 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4621 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4681 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4741 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4801 tcc
//
