LOCUS       sequence-14347-e        6137 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 6137)
  AUTHORS   Ryffel GU, Werdien D, Turan G, Gerhards A, Goosses S, Senkel S
  TITLE     Tagging muscle cell lineages in development and tail regeneration 
            using Cre recombinase in transgenic Xenopus.
  JOURNAL   Nucleic Acids Res. 2003 Apr 15. 31(8):e44.
  PUBMED    12682379
REFERENCE   2  (bases 1 to 6137)
  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. 2003 Apr 15. 31(8):e44."
COMMENT     Sequence Label: LCMV:ECFP(loxP)(FRT)EYFP
FEATURES             Location/Qualifiers
     source          1..6137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(34..67)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    75..108
                     /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      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(124..126,127..129,130..843)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    957..1091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1099..1132)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1140..1173
                     /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      1194..1203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1200..1202,1203..1205,1206..1919)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     intron          2019..2084
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2131..2265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2419..2439)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2419..2437)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2458..2474)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2458..2474)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2471..2493)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2482..2498
                     /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(2506..2512,2513..2530,2531..2536))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2551..2572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2689..2706)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2860..3448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2940..2959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3619..4410,4411..4479))
                     /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     4242..4261
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4480..4584)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4610..5065)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4747..4768)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4959..4978
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5153..6137
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttataactt cgtatagcat acattatacg
       61 aagttataac tagtgaagtt cctattctct agaaagtata ggaacttcag atccatcgcc
      121 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      181 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      241 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      301 accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg
      361 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      421 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      481 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      541 cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag
      601 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
      661 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      721 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
      781 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
      841 taatctagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
      901 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
      961 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1021 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1081 atgtctggat ctgaattcat aacttcgtat agcatacatt atacgaagtt ataggatccg
     1141 aagttcctat tctctagaaa gtataggaac ttcccgggcc taggagatcc atcgccacca
     1201 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1261 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1321 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1381 tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     1441 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1501 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1561 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1621 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     1681 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1741 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1801 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1861 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     1921 ctagtcaggg gggatctgca tggatctttg tgaaggaacc ttacttctgt ggtgtgacat
     1981 aattggacaa actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg
     2041 tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcca acctatggaa
     2101 ctgatgaatg ggagcagtgg tggaatgcca gatccagaca tgataagata cattgatgag
     2161 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2221 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2281 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     2341 ctctacaaat gtggtatggc tgattatgat ctgcggccgc ggcgccaatg cattgggccc
     2401 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     2461 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2521 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2581 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2641 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2701 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2761 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2821 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2881 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2941 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3001 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3061 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3121 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3181 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3241 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3301 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3361 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3421 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3481 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3541 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3601 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3661 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3721 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3781 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3841 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3901 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3961 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4021 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4081 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4141 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4201 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4261 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4321 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4381 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4441 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4501 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4561 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     4621 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4681 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4741 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4801 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4861 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4921 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4981 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5041 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     5101 gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ggcggccgcg tcgaccatag
     5161 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     5221 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     5281 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggact tggcactgtg
     5341 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     5401 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     5461 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     5521 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     5581 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     5641 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     5701 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     5761 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     5821 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     5881 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
     5941 attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg
     6001 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa
     6061 tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca
     6121 atgctcgttt agggaac
//
