LOCUS       sequence-14348-e        6046 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 6046)
  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 6046)
  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)EYFP
FEATURES             Location/Qualifiers
     source          1..6046
                     /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)."
     regulatory      78..87
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(84..86,87..89,90..803)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    917..1051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1053..1086)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1103..1112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1109..1111,1112..1114,1115..1828)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     intron          1928..1993
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2040..2174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2328..2348)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2328..2346)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2367..2383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2367..2383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2380..2402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2391..2407
                     /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(2415..2421,2422..2439,2440..2445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2460..2481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2598..2615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2769..3357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2849..2868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3528..4319,4320..4388))
                     /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     4151..4170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4389..4493)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4519..4974)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4656..4677)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4868..4887
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5062..6046
                     /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 aagttatagg atccatcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      121 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      181 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      241 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca
      301 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      361 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      421 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      481 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata
      541 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
      601 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
      661 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
      721 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
      781 gcatggacga gctgtacaag taatctagag gatcataatc agccatacca catttgtaga
      841 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
      901 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
      961 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1021 actcatcaat gtatcttatc atgtctggat ccataacttc gtatagcata cattatacga
     1081 agttatacct aggagatcca tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1141 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1201 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1261 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg
     1321 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1381 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1441 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1501 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1561 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1621 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1681 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     1741 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1801 tctcggcatg gacgagctgt acaagtaatc tagtcagggg ggatctgcat ggatctttgt
     1861 gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag
     1921 ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt
     1981 ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgccag
     2041 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2101 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2161 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2221 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     2281 tgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg
     2341 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2401 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2461 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2521 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2581 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2641 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2701 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2761 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2821 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2881 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2941 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3001 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3061 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3181 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4501 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4561 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4621 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4681 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4741 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4801 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4861 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4921 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     4981 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     5041 gccagtcgag gcggccgcgt cgaccatagc caattcaata tggcgtatat ggactcatgc
     5101 caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc
     5161 caattcaata tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc
     5221 caattcaata tggcggactt ggcactgtgc caactgggga ggggtctact tggcacggtg
     5281 ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg
     5341 ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat
     5401 ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata
     5461 tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc
     5521 atataccata tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg
     5581 gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc
     5641 gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt
     5701 caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc
     5761 tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct
     5821 attaatagta acttggcaag tacattacta ttggaagtac gccagggtac attggcagta
     5881 ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg
     5941 acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag
     6001 gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
