LOCUS       sequence-14344-e        5328 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 5328)
  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 5328)
  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:ECFP(FRT)MCS
FEATURES             Location/Qualifiers
     source          1..5328
                     /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"
     primer_bind     complement(128..149)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(389..408)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     736..757
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     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)"
     intron          1210..1275
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    1322..1456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1610..1630)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1610..1628)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1649..1665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1649..1665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1662..1684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1673..1689
                     /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(1697..1703,1704..1721,1722..1727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1742..1763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1880..1897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2051..2639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2131..2150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2810..3601,3602..3670))
                     /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     3433..3452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3671..3775)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3801..4256)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3938..3959)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4150..4169
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4344..5328
                     /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 ctcaccggtg actagtcagg ggggatctgc atggatcttt
     1141 gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa
     1201 agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt
     1261 gtttgtgtat tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc
     1321 agatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1381 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1441 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1501 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     1561 tctgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
     1621 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1681 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1741 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1861 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2221 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2461 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3781 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3841 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3901 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3961 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4021 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4081 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4141 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4201 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     4261 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4321 acgccagtcg aggcggccgc gtcgaccata gccaattcaa tatggcgtat atggactcat
     4381 gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt
     4441 gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat
     4501 gccaattcaa tatggcggac ttggcactgt gccaactggg gaggggtcta cttggcacgg
     4561 tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg
     4621 tgccaataat ggcggccata ttggctatat gccaggatca atatataggc aatatccaat
     4681 atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca
     4741 tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc
     4801 ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa
     4861 tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg
     4921 gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac
     4981 gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt
     5041 tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat
     5101 ctattaatag taacttggca agtacattac tattggaagt acgccagggt acattggcag
     5161 tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat
     5221 tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt
     5281 aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaac
//
