LOCUS       sequence-299342-        5078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXE3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5078)
  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 5078)
  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-FOXE3
FEATURES             Location/Qualifiers
     source          1..5078
                     /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    1130..1264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1154..1173)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1208..1227
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1377..1397)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1377..1395)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1416..1432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1416..1432)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1429..1451)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1440..1456
                     /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(1464..1470,1471..1488,1489..1494))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1509..1530
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1647..1664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1818..2406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1898..1917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2577..3368,3369..3437))
                     /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     3200..3219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3438..3542)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3568..4023)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3705..3726)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3917..3936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4094..5078
                     /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 tcgaattcgc ggggcgcagc gacatggatc cgcccgccgc gttctctggc
      181 ttccctgccc tgccagcggt cgcgccgtcg gggccgccgc cgtcgccgct cgcaggagcc
      241 gagccagggc gggagccaga ggaggcggcg gctggccgcg gagaggcggc ccccacgccc
      301 gcgcccggcc cggggcggcg gcggcggccc ctgcagcgcg ggaagccgcc ctactcgtac
      361 atcgcgctca tcgccatggc tctggcgcac gccccgggcc gccgcctcac gctggccgcc
      421 atctaccgct tcatcaccga acgctttgcc ttctaccgcg acagcccgcg caagtggcag
      481 aacagcatcc gccacaatct cacgctcaac gactgcttcg tcaaggtgcc ccgcgagccg
      541 ggcaacccgg gcaagggcaa ctactggacg ctggaccccg cggccgcaga catgttcgac
      601 aacggcagct tcctgcggcg ccgcaagcgc ttcaagcgcg ctgagctgcc cgcgcacgcg
      661 gccgcggcgc cagggccgcc gctccccttc ccctacgcgc cctacgcgcc cgcgcccggc
      721 cccgcgctgc tggtgccgcc gccttctgcc ggaccgggcc cctcgccgcc cgcgcgtctg
      781 ttcagcgtcg acagcctggt gaacctgcag ccggagctag cggggctggg cgcccccgag
      841 ccgccctgct gcgccgcgcc cgacgccgca gccgcagcct tcccgccctg cgctgccgcc
      901 gcctccccgc cactctactc gcaggtcccc gaccgcctgg tactgcccgc gacgcgcccc
      961 ggccccggcc cgctgcccgc tgagcccctc ctggccttgg ccgggccggc agccgctctc
     1021 ggcccgctca gccctgggga ggcctacctg aggcagccgg gcttcgcgtc ggggctggag
     1081 cgctacctgt gactcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat
     1141 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1201 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1261 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1321 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     1381 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1441 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     1501 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1561 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1621 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1681 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1741 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1801 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1861 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1921 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1981 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2041 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2101 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2161 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2221 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2281 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2341 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2401 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2461 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2521 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2581 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2641 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2701 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2761 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2821 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2881 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2941 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3001 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3061 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3121 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3181 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3241 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3301 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3361 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3421 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3481 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3541 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     3601 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3661 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3721 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3781 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3841 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3901 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3961 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4021 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4081 cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa
     4141 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
     4201 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     4261 tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     4321 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     4381 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     4441 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     4501 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     4561 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     4621 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     4681 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     4741 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
     4801 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
     4861 taacttggca agtacattac tattggaagt acgccagggt acattggcag tactcccatt
     4921 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
     4981 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
     5041 aatgggaggt ctatataagc aatgctcgtt tagggaac
//
