LOCUS       sequence-299332-        5558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXD3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5558)
  AUTHORS   Moparthi L, Koch S
  TITLE     A uniform expression library for the exploration of FOX 
            transcription factor biology.
  JOURNAL   Differentiation. 2020 Sep - Oct;115:30-36. doi: 
            10.1016/j.diff.2020.08.002. Epub 2020 Aug 20.
  PUBMED    32858261
REFERENCE   2  (bases 1 to 5558)
  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: 
            "Differentiation. 2020 Sep - Oct"; volume: "115"; pages: "30-36. 
            doi: 10.1016/j.diff.2020.08.002. Epub 2020 Aug 20"
COMMENT     Sequence Label: Flag-FOXD3
FEATURES             Location/Qualifiers
     source          1..5558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      85..94
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             94..117
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1610..1744
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1634..1653)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1688..1707
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1857..1877)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1857..1875)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1896..1912)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1896..1912)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1909..1931)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1920..1936
                     /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(1944..1950,1951..1968,1969..1974))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1989..2010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2127..2144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2298..2886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2378..2397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3057..3848,3849..3917))
                     /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     3680..3699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3918..4022)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4048..4503)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4185..4206)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4397..4416
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4574..5558
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcagccacc atggattaca aggacgacga cgataaggga
      121 tcccatcgat tcgaattcac cctctccggc ggcggcagcg ccagcgacat gtccggccag
      181 acggtgctga cggccgagga cgtggacatc gatgtggtgg gcgagggcga cgacgggctg
      241 gaagagaagg acagcgacgc aggttgcgat agccccgcgg ggccgccgga gctgcgcctg
      301 gacgaggcgg acgaggtgcc cccggcggca ccccatcacg gacagcctca gccgccccac
      361 cagcagcccc tgacattgcc caaggaggcg gccggagccg gggccggacc ggggggcgac
      421 gtgggcgcgc cggaggcgga cggctgcaag ggcggtgttg gcggcgagga gggcggcgcg
      481 agcggcggcg ggcctggcgc gggcagcggt tcggcgggag gcctggcccc gagcaagccc
      541 aagaacagcc tagtgaagcc gccttactcg tacatcgcgc tcatcaccat ggccatcctg
      601 cagagcccgc agaagaagct gaccctgagc ggcatctgcg agttcatcag caaccgcttc
      661 ccctactaca gggagaagtt ccccgcctgg cagaacagca tccgccacaa cctctcactc
      721 aacgactgct tcgtcaagat cccccgcgag ccgggcaacc cgggcaaggg caactactgg
      781 accctggacc cgcagtccga ggacatgttc gacaacggca gcttcctgcg gcgccggaaa
      841 cgcttcaagc gccaccagca ggagcacctg cgcgagcaga cggcgctcat gatgcagagc
      901 ttcggcgctt acagcctggc ggcggcggcc ggcgccgcgg gaccctacgg ccgcccctac
      961 ggcctgcacc ctgcggcggc ggccggtgcc tattcgcacc cggcagcggc ggcggccgcg
     1021 gctgctgcgg cggcgctcca gtacccgtac gcgctgccgc cggtggcacc ggtgctgcct
     1081 cccgctgtgc cgctgctgcc ctcgggcgag ctgggccgca aagcggccgc cttcggctca
     1141 cagctcggcc cgggcctgca gctgcagctc aatagcctgg gcgccgccgc ggccgctgcg
     1201 ggcacagcgg gcgccgcggg caccaccgcg tcgctcatca agtccgagcc aagcgcgcgg
     1261 ccgtcgttca gcatcgagaa catcataggt gggggccccg cggctcctgg gggctcggcg
     1321 gtgggcgctg gggtcgccgg cggcactggg ggttcagggg gcggcagcac ggcgcagtcg
     1381 tttctgcggc cacccgggac cgtgcagtcg gcagcgctca tggccaccca ccaaccgctg
     1441 tcgctgagcc ggacgactgc caccatcgcg cccattctta gcgtgccact ctccggacag
     1501 tttctgcagc ccgcagcctc ggccgccgcc gctgctgcgg ccgccgctca agccaaatgg
     1561 ccggcgcaat agctcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat
     1621 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1681 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1741 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1801 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     1861 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1921 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     1981 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2041 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2101 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2161 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2221 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2281 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2701 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2881 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3121 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3181 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3241 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3301 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3361 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3421 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3481 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3541 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3601 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3661 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3721 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3781 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3841 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3901 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3961 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4021 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     4081 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4141 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4201 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4261 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4321 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4381 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4441 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4501 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4561 cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa
     4621 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
     4681 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     4741 tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     4801 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     4861 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     4921 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     4981 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     5041 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     5101 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     5161 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     5221 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
     5281 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
     5341 taacttggca agtacattac tattggaagt acgccagggt acattggcag tactcccatt
     5401 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
     5461 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
     5521 aatgggaggt ctatataagc aatgctcgtt tagggaac
//
