LOCUS       sequence-14393-e        7376 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 7376)
  AUTHORS   Sauert K, Kahnert S, Roose M, Gull M, Brandli AW, Ryffel GU, Waldner
            C
  TITLE     Heat-shock mediated overexpression of HNF1beta mutations has 
            differential effects on gene expression in the Xenopus pronephric 
            kidney.
  JOURNAL   PLoS One. 2012;7(3):e33522. Epub 2012 Mar 15.
  PUBMED    22438943
REFERENCE   2  (bases 1 to 7376)
  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: "PLoS One.";
            date: "2012"; volume: "7(3)"; pages: "e33522. Epub 2012 Mar 15"
COMMENT     Sequence Label: pLCMV:ECFP(loxP)(FRT)HNF1B
FEATURES             Location/Qualifiers
     source          1..7376
                     /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"
     primer_bind     complement(169..190)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(430..449)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     777..798
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     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)"
     CDS             1202..1231
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1241..1270
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1280..1309
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1319..1348
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1358..1387
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1421..1450
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     intron          3258..3323
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    3370..3504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3658..3678)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3658..3676)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3697..3713)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3697..3713)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3710..3732)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3721..3737
                     /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(3745..3751,3752..3769,3770..3775))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3790..3811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3928..3945)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4099..4687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4179..4198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4858..5649,5650..5718))
                     /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     5481..5500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5719..5823)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5849..6304)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5986..6007)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6198..6217
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6392..7376
                     /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 taggatcgat ttaaagctat
     1201 ggagcaaaag ctcatttctg aagaggactt gaatgaaatg gagcaaaagc tcatttctga
     1261 agaggacttg aatgaaatgg agcaaaagct catttctgaa gaggacttga atgaaatgga
     1321 gcaaaagctc atttctgaag aggacttgaa tgaaatggag caaaagctca tttctgaaga
     1381 ggacttgaat gaaatggaga gcttgggcga cctcaccatg gagcaaaagc tcatttctga
     1441 agaggacttg aattcaagct tccatggtgt ccaagctcac gtcgctccag caagaactcc
     1501 tgagcgccct gctgagctcc ggggtcacca aggaggtgct ggttcaggcc ttggaggagt
     1561 tgctgccatc cccgaacttc ggggtgaagc tggagacgct gcccctgtcc cctggcagcg
     1621 gggccgagcc cgacaccaag ccggtcttcc atactctcac caacggccac gccaagggcc
     1681 gcttgtccgg cgacgagggc tccgaggacg gcgacgacta tgacacacct cccatcctca
     1741 aggagctgca ggcgctcaac accgaggagg cggcggagca gcgggcggag gtggaccgga
     1801 tgctcagtga ggacccttgg agggctgcta aaatgatcaa gggttacatg cagcaacaca
     1861 acatccccca gagggaggtg gtcgatgtca ccggcctgaa ccagtcgcac ctctcccagc
     1921 atctcaacaa gggcacccct atgaagaccc agaagcgtgc cgctctgtac acctggtacg
     1981 tcagaaagca acgagagatc ctccgacaat tcaaccagac agtccagagt tctggaaata
     2041 tgacagacaa aagcagtcag gatcagctgc tgtttctctt tccagagttc agtcaacaga
     2101 gccatgggcc tgggcagtcc gatgatgcct gctctgagcc caccaacaag aagatgcgcc
     2161 gcaaccggtt caaatggggg cccgcgtccc agcaaatctt gtaccaggcc tacgatcggc
     2221 aaaagaaccc cagcaaggaa gagagagagg ccttagtgga ggaatgcaac agggcagaat
     2281 gtttgcagcg aggggtgtcc ccctccaaag cccacggcct gggctccaac ttggtcactg
     2341 aggtccgtgt ctacaactgg tttgcaaacc gcaggaagga ggaggcattc cggcaaaagc
     2401 tggccatgga cgcctatagc tccaaccaga ctcacagcct gaacgctctc ccacggctcc
     2461 ccccaccacc agcccagctc ctctcctcca aacaagctgt caggagtgcg ctacagccag
     2521 cagggaaaca atgagatcac ttcctcctca acaatcagtc accatggcaa cagcgccatg
     2581 gtgaccagcc agtcggtttt acagcaagtc tccccagcca gcctggaccc aggccacaat
     2641 ctcctctcac ctgatggtaa aatgatctca gtctcaggag gaggtttgcc cccagtcagc
     2701 accttgacga atatccacag cctctcccac cataatcccc agcaatctca aaacctcatc
     2761 atgacacccc tctctggagt catggcaatt gcacaaagcc tcaacacctc ccaagcacag
     2821 agtgtccctg tcatcaacag tgtggccggc agcctggcag ccctgcagcc cgtccagttc
     2881 tcccagcagc tgcacagccc tcaccagcag cccctcatgc agcagagccc aggcagccac
     2941 atggcccagc agcccttcat ggcagctgtg actcagctgc agaactcaca catgtacgca
     3001 cacaagcagg aaccccccca gtattcccac acctcccggt ttccatctgc aatggtggtc
     3061 acagatacca gcagcatcag tacactcacc aacatgtctt caagtaaaca gtgtcctcta
     3121 caagcctggt gacggacgtc taggagatct caccggtgac tagtcagggg ggatctgcat
     3181 ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag
     3241 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga
     3301 ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt
     3361 ggaatgccag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3421 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3481 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3541 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     3601 gattatgatc tgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     3661 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3721 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3781 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3841 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3901 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3961 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4021 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4261 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4501 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5701 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5761 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5821 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     5881 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5941 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     6001 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     6061 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     6121 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     6181 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     6241 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     6301 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6361 tcgctattac gccagtcgag gcggccgcgt cgaccatagc caattcaata tggcgtatat
     6421 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
     6481 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
     6541 ggcaccatgc caattcaata tggcggactt ggcactgtgc caactgggga ggggtctact
     6601 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat
     6661 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa
     6721 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc
     6781 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat
     6841 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt
     6901 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga
     6961 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc
     7021 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc
     7081 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac
     7141 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac gccagggtac
     7201 attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac
     7261 atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa
     7321 tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
