LOCUS       sequence-14289-e        6203 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 6203)
  AUTHORS   Werdien D, Peiler G, Ryffel GU
  TITLE     FLP and Cre recombinase function in Xenopus embryos.
  JOURNAL   Nucleic Acids Res. 2001 Jun 1. 29(11):E53-3.
  PUBMED    11376165
REFERENCE   2  (bases 1 to 6203)
  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: "Nucleic 
            Acids Res. 2001 Jun 1. 29(11):E53-3."
COMMENT     Sequence Label: pCMV:DsRed(FRT)GFP
FEATURES             Location/Qualifiers
     source          1..6203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    174..207
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        215..233
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      272..281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(278..280,281..283,284..958)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(461..481)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     679..698
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1089..1223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1225..1258
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             1285..2001
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1315..1343)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1938..1958
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          2099..2164
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2211..2345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2499..2519)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2499..2517)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2538..2554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2538..2554)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2551..2573)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2562..2578
                     /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(join(2586..2592,2593..2610,2611..2616))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2631..2652
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2769..2786)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2940..3528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3020..3039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3699..4490,4491..4559))
                     /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     4322..4341
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4560..4664)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4690..5145)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4827..4848)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5039..5058
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5219..6203
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccatcgat tcccgggaat tcaggcctct cgagcctcta
      121 ggaacagtcg acgacactgc agagacctac ttcactaaca accggtacag ttcgaagttc
      181 ctattctcta gaaagtatag gaacttcaag cttgatttag gtgacactat agaatacaag
      241 ctacttgttc tttttgcagg atccaccggt cgccaccatg gtgcgctcct ccaagaacgt
      301 catcaaggag ttcatgcgct tcaaggtgcg catggagggc accgtgaacg gccacgagtt
      361 cgagatcgag ggcgagggcg agggccgccc ctacgagggc cacaacaccg tgaagctgaa
      421 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta
      481 cggctccaag gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt
      541 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
      601 gacccaggac tcctccctgc aggacggctg cttcatctac aaggtgaagt tcatcggcgt
      661 gaacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac
      721 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccacaagg ccctgaagct
      781 gaaggacggc ggccactacc tggtggagtt caagtccatc tacatggcca agaagcccgt
      841 gcagctgccc ggctactact acgtggactc caagctggac atcacctccc acaacgagga
      901 ctacaccatc gtggagcagt acgagcgcac cgagggccgc caccacctgt tcctgtagcg
      961 gccactcgag catgctctag aggatcataa tcagccatac cacatttgta gaggttttac
     1021 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     1081 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1141 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1201 atgtatctta tcatgtctgg atccgaagtt cctattctct agaaagtata ggaacttccc
     1261 gggcctagga gatccacagc caccatgagt aaaggagaag aacttttcac tggagttgtc
     1321 ccaattcttg ttgaattaga tggtgatgtt aatgggtaca aattttctgt cagtggagag
     1381 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
     1441 ctacctgttc catggccaac acttgtcact actctcactt atggtgttca atgcttttca
     1501 agatacccag atcatatgaa gcggcacgac ttcttcaaga gcgccatgcc tgagggatac
     1561 gtgcaggaga ggaccatctt cttcaaggac gacgggaact acaagacacg tgctgaagtc
     1621 aagtttgagg gagacaccct cgtcaacagg atcgagctta agggaatcga tttcaaggag
     1681 gacggaaaca tcctcggcca caagttggaa tacaactaca actcccacaa cgtatacatc
     1741 atggccgaca agcagaagaa cggcatcaaa gccaacttca agacccgcca caacatcgaa
     1801 gacggcggcg tgcaactcgc tgatcattat caacaaaata ctccaattgg cgatgaccct
     1861 gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac
     1921 gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc
     1981 atggatgaac tatacaaata actcgagcct ctagtcaggg gggatctttg tgaaggaacc
     2041 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt
     2101 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt
     2161 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcca gatccagaca
     2221 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2281 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2341 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2401 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat ctgcggccgc
     2461 ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     2521 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2581 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2641 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2701 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2761 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2821 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2881 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3121 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3181 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3241 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3301 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3361 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3421 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3601 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3661 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3721 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3781 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3841 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3901 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3961 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4021 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4081 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4141 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4201 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4261 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4321 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4381 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4441 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4501 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4561 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4621 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4681 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4741 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4801 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4861 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4921 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4981 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5041 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5101 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     5161 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga
     5221 ccatagccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga
     5281 cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga
     5341 ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacttggc
     5401 actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg
     5461 ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc
     5521 tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt
     5581 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
     5641 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
     5701 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
     5761 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
     5821 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
     5881 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
     5941 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
     6001 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
     6061 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
     6121 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
     6181 taagcaatgc tcgtttaggg aac
//
