LOCUS       sequence-62738-e        5640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5640)
  AUTHORS   Oliveira AF, Yasuda R
  TITLE     An improved Ras sensor for highly sensitive and quantitative 
            FRET-FLIM imaging.
  JOURNAL   PLoS One. 2013;8(1):e52874. doi: 10.1371/journal.pone.0052874. Epub 
            2013 Jan 14.
  PUBMED    23349692
REFERENCE   2  (bases 1 to 5640)
  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"; journalName: "PLoS One.";
            date: "2013"; volume: "8(1):e52874. doi"; pages: " 
            10.1371/journal.pone.0052874. Epub 2013 Jan 14"
COMMENT     Sequence Label: FRas2-F Acceptor
FEATURES             Location/Qualifiers
     source          1..5640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     718..742
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1072..1081
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1078..1752
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1078..1752
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2053..2730
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2053..2730
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     polyA_signal    2754..2875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2765..2784)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2819..2838
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3056..3511
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3143..3162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3353..3374
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3578..3597)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3697..3719
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3757..3775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3843..3947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3948..4016,4017..4808)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4166..4185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4979..5567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5468..5487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt acttaatacg actcactata ggctagcctc gagaattcgt cgccaccatg
     1081 gcctcctccg aggacgtcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc
     1141 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     1201 cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg
     1261 tcccctcagt tccagtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     1321 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     1381 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     1441 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     1501 ggctgggagg cctccaccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     1561 aagatgaggc tgaagctgaa ggacggcggc cactacgacg ccgaggtcaa gaccacctac
     1621 atggccaaga agcccgtgca gctgcccggc gcctacaaga ccgacatcaa gctggacatc
     1681 acctcccaca acgaggacta caccatcgtg gaacagtacg agcgcgccga gggccgccac
     1741 tccaccggcg cctacagatc taccatggat ccttctaaga caagcaacac tatcgctgtt
     1801 ttcttgccga acaagcaaag aacagtggtc aatgtgcgaa atggaatgag cttgcatgac
     1861 tgccttatga aagcactcaa ggtgaggggc ctgcaaccag agtgctgtgc agtgttcaga
     1921 cttctccacg aacacaaagg tgcaaaagca cgcttagatt ggaatactga tgctgcgtct
     1981 ttgattggag aagaacttca agtagatttc ctgggaattc tgcagtcgac ggtaccgcgg
     2041 gcccgggatc caatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg
     2101 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2161 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
     2221 gcctgggaca tcctgtcccc tcagttccag tacggctcca aggcctacgt gaagcacccc
     2281 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2341 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2401 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2461 cagaagaaga ccatgggctg ggaggcctcc accgagcgga tgtaccccga ggacggcgcc
     2521 ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag
     2581 gtcaagacca cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac
     2641 atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     2701 gccgagggcc gccactccac cggcgcctaa gcggccgctt cgagcagaca tgataagata
     2761 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2821 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2881 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     2941 caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgggctggc gtaatagcga
     3001 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     3061 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3121 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3181 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3241 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3301 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3361 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3421 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3481 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     3541 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     3601 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3721 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3781 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5101 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct
//
