LOCUS       sequence-14394-e        7383 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 7383)
  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 7383)
  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_A263ins
FEATURES             Location/Qualifiers
     source          1..7383
                     /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          3265..3330
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    3377..3511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3665..3685)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3665..3683)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3704..3720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3704..3720)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3717..3739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3728..3744
                     /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(3752..3758,3759..3776,3777..3782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3797..3818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3935..3952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4106..4694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4186..4205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4865..5656,5657..5725))
                     /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     5488..5507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5726..5830)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5856..6311)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5993..6014)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6205..6224
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6399..7383
                     /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 ggccttagtg gaggaatgca acagggcaga
     2281 atgtttgcag cgaggggtgt ccccctccaa agcccacggc ctgggctcca acttggtcac
     2341 tgaggtccgt gtctacaact ggtttgcaaa ccgcaggaag gaggaggcat tccggcaaaa
     2401 gctggccatg gacgcctata gctccaacca gactcacagc ctgaaccctc tgctctccca
     2461 cggctccccc caccaccagc ccagctcctc tcctccaaac aagctgtcag gagtgcgcta
     2521 cagccagcag ggaaacaatg agatcacttc ctcctcaaca atcagtcacc atggcaacag
     2581 cgccatggtg accagccagt cggttttaca gcaagtctcc ccagccagcc tggacccagg
     2641 ccacaatctc ctctcacctg atggtaaaat gatctcagtc tcaggaggag gtttgccccc
     2701 agtcagcacc ttgacgaata tccacagcct ctcccaccat aatccccagc aatctcaaaa
     2761 cctcatcatg acacccctct ctggagtcat ggcaattgca caaagcctca acacctccca
     2821 agcacagagt gtccctgtca tcaacagtgt ggccggcagc ctggcagccc tgcagcccgt
     2881 ccagttctcc cagcagctgc acagccctca ccagcagccc ctcatgcagc agagcccagg
     2941 cagccacatg gcccagcagc ccttcatggc agctgtgact cagctgcaga actcacacat
     3001 gtacgcacac aagcaggaac ccccccagta ttcccacacc tcccggtttc catctgcaat
     3061 ggtggtcaca gataccagca gcatcagtac actcaccaac atgtcttcaa gtaaacagtg
     3121 tcctctacaa gcctggtgac ggacgtctag gagatctcac cggtgactag tcagggggga
     3181 tctgcatgga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta
     3241 cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa
     3301 ctactgattc taattgtttg tgtattttag attccaacct atggaactga tgaatgggag
     3361 cagtggtgga atgccagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3421 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3481 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3541 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3601 tatggctgat tatgatctgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt
     3661 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3721 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3781 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3841 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3901 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4741 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4801 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4861 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4921 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4981 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5041 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5101 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5161 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5221 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5281 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5341 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5401 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5461 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5521 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5581 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5641 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5701 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5821 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     5881 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5941 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     6001 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     6061 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     6121 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     6181 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     6241 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     6301 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6361 ggcctcttcg ctattacgcc agtcgaggcg gccgcgtcga ccatagccaa ttcaatatgg
     6421 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
     6481 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
     6541 cggacttggc accatgccaa ttcaatatgg cggacttggc actgtgccaa ctggggaggg
     6601 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
     6661 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
     6721 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
     6781 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
     6841 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
     6901 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
     6961 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
     7021 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
     7081 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
     7141 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
     7201 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
     7261 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
     7321 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
     7381 aac
//
