LOCUS       sequence-299553-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  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 6344)
  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-FOXP2
FEATURES             Location/Qualifiers
     source          1..6344
                     /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    2396..2530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2420..2439)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2474..2493
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2643..2663)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2643..2661)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2682..2698)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2682..2698)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2695..2717)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2706..2722
                     /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(2730..2736,2737..2754,2755..2760))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2775..2796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2913..2930)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3084..3672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3164..3183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3843..4634,4635..4703))
                     /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     4466..4485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4704..4808)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4834..5289)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4971..4992)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5183..5202
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5360..6344
                     /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 tcgaattcat gcaggaatct gcgacagaga caataagcaa cagttcaatg
      181 aatcaaaatg gaatgagcac tctaagcagc caattagatg ctggcagcag agatggaaga
      241 tcaagtggtg acaccagctc tgaagtaagc acagtagaac tgctgcatct gcaacaacag
      301 caggctctcc aggcagcaag acaacttctt ttacagcagc aaacaagtgg attgaaatct
      361 cctaagagca gtgataaaca gagaccactg caggaattgc ttccagaaac aaaattatgt
      421 atctgtggcc actcttctgg tgatgggcat cctcacaaca catttgcagt gcctgtgtca
      481 gtggccatga tgactcccca ggtgatcacc cctcagcaaa tgcagcagat ccttcagcaa
      541 caagtcctgt ctcctcagca gctacaagcc cttctccaac aacagcaggc tgtcatgctg
      601 cagcagcaac aactacaaga gttttacaag aaacagcaag agcagttaca tcttcagctt
      661 ttgcagcagc agcagcaaca gcagcagcag caacaacagc agcaacaaca gcagcagcaa
      721 caacaacaac aacagcagca acaacagcag cagcagcagc aacagcagca gcagcagcaa
      781 cagcatcctg gaaagcaagc gaaagagcag cagcagcagc agcagcagca acagcaattg
      841 gcagcccagc agcttgtctt ccagcagcag cttctccaga tgcaacaact ccagcagcag
      901 cagcatctgc tcagccttca gcgtcaggga ctcatctcca ttccacctgg ccaggcagca
      961 cttcctgtcc aatcgctgcc tcaagctggc ttaagtcctg ctgagattca gcagttatgg
     1021 aaagaagtga ctggagttca cagtatggaa gacaatggca ttaaacatgg agggctagac
     1081 ctcactacta acaattcctc ctcgactacc tcctccaaca cttccaaagc atcaccacca
     1141 ataactcatc attccatagt gaatggacag tcttcagttc taagtgcaag acgagacagc
     1201 tcgtcacatg aggagactgg ggcctctcac actctctatg gccatggagt ttgcaaatgg
     1261 ccaggctgtg aaagcatttg tgaagatttt ggacagtttt taaagcacct taacaatgaa
     1321 cacgcattgg atgaccgaag cactgctcag tgtcgagtgc aaatgcaggt ggtgcaacag
     1381 ttagaaatac agctttctaa agaacgcgaa cgtcttcaag caatgatgac ccacttgcac
     1441 atgcgaccct cagagcccaa accatctccc aaacctctaa atctggtgtc tagtgtcacc
     1501 atgtcgaaga atatgttgga gacatcccca cagagcttac ctcaaacccc taccacacca
     1561 acggccccag tcaccccgat tacccaggga ccctcagtaa tcaccccagc cagtgtgccc
     1621 aatgtgggag ccatacgaag gcgacattca gacaaataca acattcccat gtcatcagaa
     1681 attgccccaa actatgaatt ttataaaaat gcagatgtca gacctccatt tacttatgca
     1741 actctcataa ggcaggctat catggagtca tctgacaggc agttaacact taatgaaatt
     1801 tacagctggt ttacacggac atttgcttac ttcaggcgta atgcagcaac ttggaagaat
     1861 gcagtacgtc ataatcttag cctgcacaag tgttttgttc gagtagaaaa tgttaaagga
     1921 gcagtatgga ctgtggatga agtagaatac cagaagcgaa ggtcacaaaa gataacagga
     1981 agtccaacct tagtaaaaaa tatacctacc agtttaggct atggagcagc tcttaatgcc
     2041 agtttgcagg ctgccttggc agagagcagt ttacctttgc taagtaatcc tggactgata
     2101 aataatgcat ccagtggcct actgcaggcc gtccacgaag acctcaatgg ttctctggat
     2161 cacattgaca gcaatggaaa cagtagtccg ggctgctcac ctcagccgca catacattca
     2221 atccacgtca aggaagagcc agtgattgca gaggatgaag actgcccaat gtccttagtg
     2281 acaacagcta atcacagtcc agaattagaa gacgacagag agattgaaga agagccttta
     2341 tctgaagatc tggaatgact cgagcctcta gaactatagt gagtcgtatt acgtagatcc
     2401 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2461 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2521 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2581 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     2641 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     2701 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     2761 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2821 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2881 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3001 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3481 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3721 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3781 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3841 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3901 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4021 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4261 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4501 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4681 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4741 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4801 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     4861 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4921 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4981 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5041 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     5101 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5161 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5221 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5281 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     5341 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
     5401 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
     5461 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
     5521 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
     5581 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
     5641 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
     5701 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
     5761 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
     5821 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
     5881 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
     5941 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
     6001 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
     6061 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
     6121 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
     6181 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
     6241 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
     6301 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaac
//
