LOCUS       sequence-299510-        5975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXJ3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5975)
  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 5975)
  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-FOXJ3
FEATURES             Location/Qualifiers
     source          1..5975
                     /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    2027..2161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2051..2070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2105..2124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2274..2294)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2274..2292)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2313..2329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2313..2329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2326..2348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2337..2353
                     /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(2361..2367,2368..2385,2386..2391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2406..2427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2544..2561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2715..3303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2795..2814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3474..4265,4266..4334))
                     /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     4097..4116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4335..4439)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4465..4920)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4602..4623)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4814..4833
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4991..5975
                     /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 tccggtttgt atggacaggc ttgtccatct gtaacttcat taaggatgac atctgaactg
      181 gagagcagcc taacgtctat ggactggtta ccacagctca ccatgagagc agccatccaa
      241 aaatctgatg ctacacaaaa tgcacatgga acaggaattt ctaagaagaa tgcactcctt
      301 gacccaaata caactctgga ccaggaagaa gtccaacagc acaaagatgg gaaacctcca
      361 tacagttatg ccagcctcat tacatttgca attaatagct cacccaaaaa gaaaatgact
      421 ttaagtgaaa tttatcagtg gatttgtgat aacttcccat attatagaga ggctggcagt
      481 ggttggaaga attccatacg acataatctg tcattgaaca aatgtttcct taaagtgcct
      541 cgatctaagg atgaccctgg aaaggggtcc tactgggcaa tagacaccaa tccgaaggaa
      601 gatgtgctgc ctactcggcc aaagaagagg gcacgatctg tagaacgggc ctcaactcca
      661 tatagcatag attcagattc tttgggaatg gagtgtatta tttcgggaag tgcctctcca
      721 actctggcaa tcaacactgt gactaacaaa gtaacattgt ataacactga tcaggatggt
      781 agtgatagcc cacgcagtag ccttaacaac agtctctcag accagagttt ggcatctgtt
      841 aatttgaaca gtgttggaag tgtgcatagt tatacaccgg tgacaagcca tccagaatca
      901 gtctctcaat cattaactcc tcagcagcaa ccacagtaca accttccaga aagagacaag
      961 caactacttt tctcagaata taattttgaa gatcttagtg cctcatttcg gagcctttat
     1021 aagtcagttt ttgagcagtc acttagtcaa caaggtttga tgaacatccc ttctgaatct
     1081 tcccagcagt cccacacttc atgtacctat cagcactctc ccagcagtac agtgagcact
     1141 cacccacaca gcaaccaaag cagcctgtcc aacagtcatg gcagtggcct caacaccaca
     1201 ggcagtaatt cggttgcaca ggtctcactg tctcaccccc agatgcacac acagccatct
     1261 ccacatcctc cccatcgacc gcatggttta ccgcagcatc cgcagcgttc cccacaccca
     1321 gcaccacacc cacagcaaca cagccagctc cagtcccctc acccccagca tccctctcca
     1381 catcaacaca tacagcacca tccgaaccat cagcatcaga cgttaacaca tcaggcaccc
     1441 ccacccccac aacaggtatc ctgtaattct ggtgtttcaa atgattggta tgcgacactt
     1501 gatatgctaa aagaaagctg tcgaattgcc agcagtgtta attggtcaga tgtagacctt
     1561 tcacagtttc aaggtctgat ggagagtatg agacaggcag atctcaagaa ctggtcttta
     1621 gaccaggttc agtttgccga tctttgttct tctcttaatc agttctttac acaaactggc
     1681 cttatacatt cacagagtaa tgttcaacaa aatgtttgtc atggtgccat gcatccaaca
     1741 aaaccttccc aacacattgg aacaggaaat ttgtacatag attctaggca aaatctccct
     1801 ccttcagtga tgccaccccc tggttatcct catatcccac aggcactcag cactccagga
     1861 acaacgatgg caggccatca cagagccatg aaccagcagc acatgatgcc ttcccaagcc
     1921 ttccagatgc ggcgttccct gcctccagat gacatccagg atgactttga ttgggattca
     1981 attgtgtagc tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat
     2041 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2101 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2161 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2221 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
     2281 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2341 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
     2401 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2461 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2521 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2581 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2641 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2701 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2761 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2821 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2881 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2941 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3001 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3061 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3121 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3181 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3241 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3301 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3361 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3421 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3721 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3901 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4021 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4141 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4201 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4321 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4381 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4441 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     4501 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4561 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4621 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4681 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4741 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4801 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4861 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4921 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4981 tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
     5041 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
     5101 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
     5161 ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
     5221 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
     5281 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
     5341 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
     5401 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
     5461 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
     5521 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
     5581 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
     5641 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
     5701 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
     5761 cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac
     5821 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
     5881 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     5941 gggaggtcta tataagcaat gctcgtttag ggaac
//
