LOCUS       Exported                8874 bp ds-DNA     circular SYN 25-DEC-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pIRES-split-KRas-S17N
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8874)
  AUTHORS   gene
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 2, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8874
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        138..517
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          890..1022
                     /note="chimeric intron"
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1067..1085
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1091..1807
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al.,
                     2004)"
                     /label=enhanced monomeric variant of CFP (Rizzo et al
                     /note="mCerulean"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1832..2101
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FKBP-rapamycin binding domain of human FRAP
                     /note="FRB"
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     misc_feature    2248..2821
                     /label=IRES
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3428..3748
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=human FK506-binding protein FKBP12
                     /note="FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3773..4486
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=Venus YFP with monomerizing A206K mutation (Nag
                     /note="mVenus"
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     promoter        complement(4543..4560)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    4578..4699
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      4885..5340
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5457..5814
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      5665..5800
                     /label=SV40 ori
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             5865..6659
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=aph(3')-II (or nptII)
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6723..6771
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
     promoter        7077..7181
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     CDS             7182..8042
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8213..8801
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1141 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1201 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1261 ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcgccc gctaccccga
     1321 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1381 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1441 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1501 cctggggcac aagctggagt acaacgccat cagcgacaac gtctatatca ccgccgacaa
     1561 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcagcgt
     1621 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1681 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     1741 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1801 gtacaagggc ggtggctcat ctggcggagg tgagatgtgg catgaaggcc tggaagaggc
     1861 atctcgtttg tactttgggg aaaggaacgt gaaaggcatg tttgaggtgc tggagccctt
     1921 gcatgctatg atggaacggg gcccccagac tctgaaggaa acatccttta atcaggccta
     1981 tggtcgagat ttaatggagg cccaagagtg gtgcaggaag tacatgaaat cagggaatgt
     2041 caaggacctc acccaagcct gggacctcta ttatcatgtg ttccgacgaa tctcaaagca
     2101 gcagatctcg tacgcgtccc ggggcggtgg ctcatctggc ggaggtgagc tcatgactga
     2161 atataaactt gtggtagttg gagctggtta agaattcacg cgtcgagcat gcatctaggg
     2221 cggccaattc cgcccctctc cccccccccc ctctccctcc ccccccccta acgttactgg
     2281 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     2341 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     2401 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     2461 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     2521 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     2581 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     2641 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     2701 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     2761 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     2821 agcttgccac aacccgggat cctctagaat gggcgtaggc aagaatgcct tgacgataca
     2881 gctaattcag aatcattttg tggacgaata tgatccaaca atagaggatt cctacaggaa
     2941 gcaagtagta attgatggag aaacctgtct cttggatatt ctcgacacag caggtcaaga
     3001 ggagtacagt gcaatgaggg accagtacat gaggactggg gagggctttc tttgtgtatt
     3061 tgccataaat aatactaaat catttgaaga tattcaccat tatagagaac aaattaaaag
     3121 agttaaggac tctgaagatg tacctatggt cctagtagga aataaatgtg atttgccttc
     3181 cagaacagta gacacaaaac aggctcagga cttagcaaga agttatggaa ttccttttat
     3241 tgaaacatca gcaaagacaa gacagggtgt tgatgatgcc ttctatacat tagttcgaga
     3301 aattcgaaaa cataaagaaa agatgagcaa agatggtggg gatcccaatt gggagctcgt
     3361 gtacacggcg cgcctgcagg gaggtggctc atctggcgga ggtcagatct cgtacgcgtc
     3421 ccggggcgga gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg
     3481 cggccagacc tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga aatttgattc
     3541 ctcccgggac agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg
     3601 ctgggaagaa ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc
     3661 agattatgcc tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt
     3721 cttcgatgtg gagcttctaa aactggaagg cggtggctca tctggcggag gtgtgagcaa
     3781 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3841 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3901 cctgaagctc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3961 cctcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     4021 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4081 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4141 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4201 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     4261 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     4321 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     4381 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4441 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagaaaa aaaagaagaa
     4501 aaagagcaag accaaatgcg tgattatgta agcggccgct tccctttagt gagggttaat
     4561 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4621 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4681 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4741 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatccgata
     4801 aggatcgatc cgggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     4861 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4921 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4981 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5041 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5101 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5161 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5221 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5281 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5341 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc
     5401 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     5461 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5521 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5581 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5641 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5701 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5761 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     5821 gattcttctg acacaacagt ctcgaactta aggctagagc caccatgatt gaacaagatg
     5881 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5941 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6001 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     6061 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6121 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6181 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6241 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6301 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6361 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     6421 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6481 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6541 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6601 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6661 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc
     6721 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     6781 cgataaggat ccgcgtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6841 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     6901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     6961 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     7021 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     7081 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     7141 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7201 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7261 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7321 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     7381 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     7441 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     7501 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     7561 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     7621 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     7681 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     7741 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     7801 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     7861 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7921 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7981 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     8041 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     8101 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     8161 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     8221 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     8281 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     8341 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     8401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     8461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     8521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     8581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     8641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     8701 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     8761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     8821 ttttacggtt cctggccttt tgctggcctt ttgctcacat ggctcgacag atct
//
