LOCUS       sequence-14375-e        6216 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 6216)
  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 6216)
  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:ECFP(FRT)EYFP
FEATURES             Location/Qualifiers
     source          1..6216
                     /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      268..277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(274..276,277..279,280..993)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1107..1241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1243..1276
                     /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      1297..1306
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1303..1305,1306..1308,1309..2022)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     intron          2112..2177
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2224..2358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2512..2532)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2512..2530)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2551..2567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2551..2567)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2564..2586)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2575..2591
                     /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(2599..2605,2606..2623,2624..2629))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2644..2665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2782..2799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2953..3541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3033..3052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3712..4503,4504..4572))
                     /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     4335..4354
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4573..4677)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4703..5158)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4840..4861)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5052..5071
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5232..6216
                     /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 atccatcgcc accatggtga gcaagggcga ggagctgttc
      301 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      361 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      421 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg
      481 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      541 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      601 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      661 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac
      721 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
      781 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
      841 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
      901 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      961 atcactctcg gcatggacga gctgtacaag taatctagag gatcataatc agccatacca
     1021 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1081 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1201 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgaagttcc tattctctag
     1261 aaagtatagg aacttcccgg gcctaggaga tccatcgcca ccatggtgag caagggcgag
     1321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1381 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
     1501 tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1561 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1741 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1861 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1921 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1981 gccgccggga tcactctcgg catggacgag ctgtacaagt aatctagtca ggggggatct
     2041 ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct acagagattt
     2101 aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa
     2161 ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg
     2221 ccagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2281 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2341 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2401 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     2461 gatctgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     2521 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2581 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2641 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2701 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2761 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2821 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2881 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3121 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3361 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     3421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3601 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3841 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4561 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4621 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4681 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     4741 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4801 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4861 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4921 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4981 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5041 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5101 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     5161 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5221 ttacgccagt cgaccatagc caattcaata tggcgtatat ggactcatgc caattcaata
     5281 tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata
     5341 tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata
     5401 tggcggactt ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga
     5461 ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg
     5521 cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc
     5581 caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca
     5641 tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata
     5701 tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat
     5761 atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca
     5821 ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc
     5881 acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc
     5941 tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta
     6001 acttggcaag tacattacta ttggaagtac gccagggtac attggcagta ctcccattga
     6061 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
     6121 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
     6181 tgggaggtct atataagcaa tgctcgttta gggaac
//
