LOCUS       sequence-299554-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXP3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  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 5418)
  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-FOXP3
FEATURES             Location/Qualifiers
     source          1..5418
                     /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    1470..1604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1494..1513)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1548..1567
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1717..1737)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1717..1735)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1756..1772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1756..1772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1769..1791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1780..1796
                     /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(1804..1810,1811..1828,1829..1834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1849..1870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1987..2004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2158..2746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2238..2257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2917..3708,3709..3777))
                     /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     3540..3559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3778..3882)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3908..4363)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4045..4066)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4257..4276
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4434..5418
                     /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 tcgaattccc caaccccagg cctggcaagc cctcggcccc ttccttggcc
      181 cttggcccat ccccaggagc ctcgcccagc tggagggctg cacccaaagc ctcagacctg
      241 ctgggggccc ggggcccagg gggaaccttc cagggccgag atcttcgagg cggggcccat
      301 gcctcctctt cttccttgaa ccccatgcca ccatcgcagc tgcagctgcc cacactgccc
      361 ctagtcatgg tggcaccctc cggggcacgg ctgggcccct tgccccactt acaggcactc
      421 ctccaggaca ggccacattt catgcaccag ctctcaacgg tggatgccca cgcccggacc
      481 cctgtgctgc aggtgcaccc cctggagagc ccagccatga tcagcctcac accacccacc
      541 accgccactg gggtcttctc cctcaaggcc cggcctggcc tcccacctgg gatcaacgtg
      601 gccagcctgg aatgggtgtc cagggagccg gcactgctct gcaccttccc aaatcccagt
      661 gcacccagga aggacagcac cctttcggct gtgccccaga gctcctaccc actgctggca
      721 aatggtgtct gcaagtggcc cggatgtgag aaggtcttcg aagagccaga ggacttcctc
      781 aagcactgcc aggcggacca tcttctggat gagaagggca gggcacaatg tctcctccag
      841 agagagatgg tacagtctct ggagcagcag ctggtgctgg agaaggagaa gctgagtgcc
      901 atgcaggccc acctggctgg gaaaatggca ctgaccaagg cttcatctgt ggcatcatcc
      961 gacaagggct cctgctgcat cgtagctgct ggcagccaag gccctgtcgt cccagcctgg
     1021 tctggccccc gggaggcccc tgacagcctg tttgctgtcc ggaggcacct gtggggtagc
     1081 catggaaaca gcacattccc agagttcctc cacaacatgg actacttcaa gttccacaac
     1141 atgcgacccc ctttcaccta cgccacgctc atccgctggg ccatcctgga ggctccagag
     1201 aagcagcgga cactcaatga gatctaccac tggttcacac gcatgtttgc cttcttcaga
     1261 aaccatcctg ccacctggaa gaacgccatc cgccacaacc tgagtctgca caagtgcttt
     1321 gtgcgggtgg agagcgagaa gggggctgtg tggaccgtgg atgagctgga gttccgcaag
     1381 aaacggagcc agaggcccag caggtgttcc aaccctacac ctggcccctg acctcgagcc
     1441 tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     1501 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1561 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1621 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg
     1681 cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     1741 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1801 acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1861 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1921 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2341 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2581 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2641 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2701 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2761 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2821 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2881 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2941 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3001 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3061 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3121 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3181 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3241 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3301 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3361 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3421 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3481 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3541 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3601 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3661 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3721 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3781 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3841 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3901 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3961 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4021 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4081 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4141 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4201 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4261 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4321 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     4381 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc
     4441 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
     4501 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
     4561 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
     4621 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
     4681 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
     4741 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
     4801 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
     4861 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
     4921 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
     4981 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
     5041 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
     5101 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
     5161 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
     5221 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
     5281 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
     5341 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
     5401 aatgctcgtt tagggaac
//
