LOCUS       sequence-14288-e        6026 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 6026)
  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 6026)
  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: LCMV:GFP(FRT)DsRed
FEATURES             Location/Qualifiers
     source          1..6026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    34..67
                     /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             83..799
                     /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(113..141)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     736..756
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    921..1055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1057..1090
                     /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)"
     regulatory      1115..1124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1121..1123,1124..1126,1127..1801)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(1304..1324)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1522..1541
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          1908..1973
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2020..2154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2308..2328)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2308..2326)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2347..2363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2347..2363)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2360..2382)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2371..2387
                     /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(2395..2401,2402..2419,2420..2425))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2440..2461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2578..2595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2749..3337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2829..2848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3508..4299,4300..4368))
                     /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     4131..4150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4369..4473)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4499..4954)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4636..4657)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4848..4867
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5042..6026
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgaagttc ctattctcta gaaagtatag
       61 gaacttcgga tccacagcca ccatgagtaa aggagaagaa cttttcactg gagttgtccc
      121 aattcttgtt gaattagatg gtgatgttaa tgggtacaaa ttttctgtca gtggagaggg
      181 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
      241 acctgttcca tggccaacac ttgtcactac tctcacttat ggtgttcaat gcttttcaag
      301 atacccagat catatgaagc ggcacgactt cttcaagagc gccatgcctg agggatacgt
      361 gcaggagagg accatcttct tcaaggacga cgggaactac aagacacgtg ctgaagtcaa
      421 gtttgaggga gacaccctcg tcaacaggat cgagcttaag ggaatcgatt tcaaggagga
      481 cggaaacatc ctcggccaca agttggaata caactacaac tcccacaacg tatacatcat
      541 ggccgacaag cagaagaacg gcatcaaagc caacttcaag acccgccaca acatcgaaga
      601 cggcggcgtg caactcgctg atcattatca acaaaatact ccaattggcg atgaccctgt
      661 ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga
      721 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
      781 ggatgaacta tacaaataac tcgagcctct agaggatcat aatcagccat accacatttg
      841 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
      901 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
      961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1021 ccaaactcat caatgtatct tatcatgtct ggatccgaag ttcctattct ctagaaagta
     1081 taggaacttc ccgggcctag gagatccacc ggtcgccacc atggtgcgct cctccaagaa
     1141 cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggcaccgtga acggccacga
     1201 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggccacaaca ccgtgaagct
     1261 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca
     1321 gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc
     1381 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     1441 cgtgacccag gactcctccc tgcaggacgg ctgcttcatc tacaaggtga agttcatcgg
     1501 cgtgaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     1561 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca aggccctgaa
     1621 gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg ccaagaagcc
     1681 cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct cccacaacga
     1741 ggactacacc atcgtggagc agtacgagcg caccgagggc cgccaccacc tgttcctgta
     1801 gcggccactc gagcatgctc tagtcagggg ggatctttgt gaaggaacct tacttctgtg
     1861 gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat
     1921 ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa
     1981 cctatggaac tgatgaatgg gagcagtggt ggaatgccag atccagacat gataagatac
     2041 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2101 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2161 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2221 aagtaaaacc tctacaaatg tggtatggct gattatgatc tgcggccgcg gcgccaatgc
     2281 attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     2341 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2401 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2461 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2521 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2581 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2641 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2701 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2761 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2821 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2881 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2941 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3001 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3061 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3121 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3181 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3241 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3301 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3361 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3421 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3481 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3721 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3781 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4021 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4141 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4201 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4381 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4441 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     4501 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4561 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4621 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4681 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4741 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4801 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4861 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4921 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     4981 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgag gcggccgcgt
     5041 cgaccatagc caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct
     5101 ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct
     5161 ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata tggcggactt
     5221 ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct
     5281 tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt
     5341 ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct
     5401 attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt
     5461 ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt
     5521 cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc
     5581 ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa
     5641 ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt
     5701 gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc
     5761 aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag
     5821 tacattacta ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc
     5881 ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa
     5941 tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct
     6001 atataagcaa tgctcgttta gggaac
//
