LOCUS       sequence-14346-e        5333 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 5333)
  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 5333)
  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)MCS
FEATURES             Location/Qualifiers
     source          1..5333
                     /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)"
     intron          1215..1280
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    1327..1461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1615..1635)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1615..1633)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1654..1670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1654..1670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1667..1689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1678..1694
                     /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(1702..1708,1709..1726,1727..1732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1747..1768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1885..1902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2056..2644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2136..2155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2815..3606,3607..3675))
                     /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     3438..3457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3676..3780)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3806..4261)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3943..3964)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4155..4174
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4349..5333
                     /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 gagatctcac cggtgactag tcagggggga tctgcatgga
     1141 tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga
     1201 tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctactgattc
     1261 taattgtttg tgtattttag attccaacct atggaactga tgaatgggag cagtggtgga
     1321 atgccagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1381 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1441 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1501 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     1561 tatgatctgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     1621 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1681 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1741 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1861 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2461 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     2521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2701 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2761 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2821 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2881 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2941 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3001 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3061 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3121 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3181 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3241 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3301 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3361 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3421 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3481 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3541 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3601 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3661 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3721 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3781 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     3841 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3901 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3961 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4021 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4081 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4141 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4201 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4261 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4321 ctattacgcc agtcgaggcg gccgcgtcga ccatagccaa ttcaatatgg cgtatatgga
     4381 ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga
     4441 ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc
     4501 accatgccaa ttcaatatgg cggacttggc actgtgccaa ctggggaggg gtctacttgg
     4561 cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg
     4621 catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat
     4681 ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta
     4741 tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg
     4801 cggtcccata taccatatat ggggcttcct aataccgccc atagccactc ccccattgac
     4861 gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt
     4921 atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca
     4981 ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg cccattcata
     5041 tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc
     5101 aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt
     5161 ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc
     5221 cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg
     5281 gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
