LOCUS       sequence-429744-        5120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXS1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5120)
  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 5120)
  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-FOXS1
FEATURES             Location/Qualifiers
     source          1..5120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..985
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1020..1038
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1020..1037
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1070..1079
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1079..1102
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2157..2291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2181..2200)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2235..2254
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2404..2424)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2404..2422)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2443..2459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2443..2459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2456..2478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2467..2483
                     /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(2491..2497,2498..2515,2516..2521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2536..2557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2674..2691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2845..3433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2925..2944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3604..4395,4396..4464))
                     /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     4227..4246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4465..4569)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4595..5050)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4732..4753)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4944..4963
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg
       61 gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg
      121 gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg
      181 gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt
      241 ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg
      301 gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta
      361 ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat atatgggttt
      421 tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc
      481 ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc
      541 tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat
      601 tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg
      661 acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca
      721 atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt
      781 acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg
      841 gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat
      901 gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta
      961 tataagcaat gctcgtttag ggaaccgcca ttctgcctgg ggacgtcgga gcaagcttga
     1021 tttaggtgac actatagaat acaagctact tgttcttttt gcaggatcag ccaccatgga
     1081 ttacaaggac gacgacgata agggatccca tcgattcgaa ttccagcagc agcctctgcc
     1141 cgggcctggc gcccccacaa ctgagccaac caagcctccc tacagctaca tcgcccttat
     1201 tgctatggcc atccagagct caccggggca gcgggccacc ctcagtggca tctaccgcta
     1261 catcatgggc cgattcgcct tctaccgcca caaccggccc ggctggcaga acagcatccg
     1321 ccacaacctg tcactcaacg agtgctttgt caaggtgccc cgcgatgacc gcaagccagg
     1381 caagggcagc tactggacgc tggaccctga ctgccacgac atgtttgagc acggcagctt
     1441 cctacgccgc cgccgccgct tcacccggca gacaggtgct gagggcaccc ggggccccgc
     1501 caaggcacgc cgtggacccc tcagggcgac cagccaggac ccaggagtcc ccaacgccac
     1561 gaccggcagg cagtgctcat tcccaccaga gctgccagat cccaagggcc taagctttgg
     1621 gggtctggtg ggggccatgc cagccagtat gtgcccagca accactgatg gcaggcctcg
     1681 gccacccatg gagcccaaag agatttccac gcccaagcct gcatgcccag gggagctccc
     1741 cgtggccacc tcatcttcct catgcccagc gtttggcttt cctgccggct tctcagaggc
     1801 tgagagtttt aataaggccc ctacgcccgt cttgtccccg gaatcaggca tcgggagcag
     1861 ctaccagtgt cggctgcagg cactgaattt ttgcatgggg gctgacccag gccttgagca
     1921 cctcttggcc tcagcagccc cctcccctgc accacccacc cctccaggct cactccgggc
     1981 cccactgccc ctgccaactg accacaagga accctgggtt gcaggtggct tccctgtcca
     2041 gggaggctcc ggctacccat tggggctgac cccctgccta taccggacgc caggaatgtt
     2101 cttctttgag taaaggcagc tcgagcctct agaactatag tgagtcgtat tacgtagatc
     2161 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2221 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2281 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     2341 gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg
     2401 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     2461 gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa
     2521 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2581 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2641 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2701 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2761 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2821 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2881 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2941 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3001 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3061 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3121 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3181 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3241 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3301 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3361 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3421 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3481 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3541 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3601 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3661 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3721 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3781 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3841 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3901 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3961 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4021 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4081 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4141 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4201 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4261 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4321 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4381 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4441 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4501 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4561 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     4621 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4681 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4741 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4801 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4861 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4921 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4981 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5041 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5101 gcctcttcgc tattacgcca
//
