LOCUS       sequence-299377-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXJ1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  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 5378)
  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-FOXJ1
FEATURES             Location/Qualifiers
     source          1..5378
                     /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    1430..1564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1454..1473)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1508..1527
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1677..1697)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1677..1695)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1716..1732)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1716..1732)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1729..1751)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1740..1756
                     /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(1764..1770,1771..1788,1789..1794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1809..1830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1947..1964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2118..2706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2198..2217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2877..3668,3669..3737))
                     /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     3500..3519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3738..3842)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3868..4323)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4005..4026)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4217..4236
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4394..5378
                     /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 tccgcggaga gctggctgcg cctctcggga gccgggccgg cggaggaggc cgggccggag
      181 ggcggcctgg aggagcccga cgccctggat gacagcctga ccagcctgca gtggctgcag
      241 gaattctcca ttctcaacgc caaggccccc gccctgcccc cggggggcac cgacccccac
      301 ggctaccacc aggtgccagg ttcagcggcg cccgggtccc ccctggcggc cgaccccgcc
      361 tgcctggggc agccacacac gccgggcaag cccacgtcgt cgtgcacgtc gcggagcgcg
      421 cccccggggc tgcaggcccc accccccgac gacgtggact acgccaccaa tccgcacgtg
      481 aagcctccct actcgtatgc cacgctcatc tgcatggcca tgcaggccag caaggccacc
      541 aagatcaccc tgtcggccat ctacaagtgg atcacggaca acttctgcta cttccgccac
      601 gcagatccca cctggcagaa ttcaatccgc cacaacctgt ctctgaacaa gtgcttcatc
      661 aaagtgcctc gggagaagga cgaaccaggc aaggggggct tctggcgcat tgacccccag
      721 tacgcggagc ggctactgag cggcgctttc aagaagcggc gactgccccc tgtccacatc
      781 cacccagcct ttgcccgcca ggccgcgcag gagcccagcg ctgtcccccg ggccgggccg
      841 ctgacggtga ataccgaggc ccagcagctg ctgcgggagt tcgaggaggc caccggggag
      901 gcgggctggg gtgcaggcga gggcaggctg gggcataagc gcaaacagcc gctgcccaag
      961 cgggtggcca aggtcccgcg gccccccagc accctgctgc ccaccccgga ggagcagggt
     1021 gagctggaac ccctcaaagg caactttgac tgggaggcca tcttcgacgc cggcactctg
     1081 ggcggggagc tgggtgcact ggaggccctg gagctgagcc cgcctctgag ccccgcctca
     1141 cacgtggacg tggacctcac catccacggc cgccacatcg actgccctgc cacctggggg
     1201 ccttcggtgg agcaggctgc cgacagcctg gacttcgatg agaccttcct ggccacatcc
     1261 ttcctgcagc acccctggga cgagagcggc agtggctgcc tgcccccgga gcccctcttt
     1321 gaggctgggg atgccaccct ggcctccgac ctgcaggact gggccagcgt gggggccttc
     1381 ttgtaagagg ccctcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat
     1441 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1501 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1561 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1621 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     1681 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1741 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     1801 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1861 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1921 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2281 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2521 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2761 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2821 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2881 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2941 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3001 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3061 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3121 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3181 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3241 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3301 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3361 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3421 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3481 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3541 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3601 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3661 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3721 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3781 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3841 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     3901 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3961 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4021 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4081 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4141 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4201 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4261 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4321 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4381 cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa
     4441 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
     4501 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     4561 tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     4621 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     4681 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     4741 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     4801 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     4861 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     4921 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     4981 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     5041 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
     5101 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
     5161 taacttggca agtacattac tattggaagt acgccagggt acattggcag tactcccatt
     5221 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
     5281 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
     5341 aatgggaggt ctatataagc aatgctcgtt tagggaac
//
