LOCUS       sequence-14350-e        6049 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 6049)
  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 6049)
  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(FRT)EYFP
FEATURES             Location/Qualifiers
     source          1..6049
                     /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)"
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(83..85,86..88,89..802)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    916..1050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1052..1085
                     /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      1106..1115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1112..1114,1115..1117,1118..1831)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     intron          1931..1996
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2043..2177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2331..2351)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2331..2349)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2370..2386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2370..2386)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2383..2405)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2394..2410
                     /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(2418..2424,2425..2442,2443..2448))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2463..2484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2601..2618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2772..3360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2852..2871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3531..4322,4323..4391))
                     /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     4154..4173
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4392..4496)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4522..4977)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4659..4680)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4871..4890
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5065..6049
                     /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 tccatcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      121 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      181 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      241 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag
      301 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      361 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      421 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      481 ggacggcaac atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat
      541 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
      601 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      661 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
      721 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
      781 catggacgag ctgtacaagt aatctagagg atcataatca gccataccac atttgtagag
      841 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
      901 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
      961 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1021 ctcatcaatg tatcttatca tgtctggatc cgaagttcct attctctaga aagtatagga
     1081 acttcccggg cctaggagat ccatcgccac catggtgagc aagggcgagg agctgttcac
     1141 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1201 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1261 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca
     1321 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1381 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1441 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1501 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1561 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1621 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1681 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     1741 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1801 cactctcggc atggacgagc tgtacaagta atctagtcag gggggatctg catggatctt
     1861 tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta
     1921 aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat
     1981 tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc
     2041 cagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2101 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2161 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2221 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     2281 atctgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     2341 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2401 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2461 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2521 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2581 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2641 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2701 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2761 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2821 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2881 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2941 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3001 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3061 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3121 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3181 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     3241 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3301 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3361 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3421 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3481 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3541 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3601 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3661 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3721 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3781 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3841 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3901 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3961 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4021 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4081 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4141 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4201 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4261 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4321 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4381 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4441 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4501 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4561 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4621 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4681 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4741 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4801 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4861 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4921 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     4981 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5041 tacgccagtc gaggcggccg cgtcgaccat agccaattca atatggcgta tatggactca
     5101 tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg
     5161 tgccaattca atatggtgga tctggacccc agccaattca atatggcgga cttggcacca
     5221 tgccaattca atatggcgga cttggcactg tgccaactgg ggaggggtct acttggcacg
     5281 gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg
     5341 gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg caatatccaa
     5401 tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc
     5461 atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt
     5521 cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca
     5581 atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat
     5641 ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga
     5701 cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg
     5761 ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata
     5821 tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca
     5881 gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca
     5941 ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg
     6001 taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
