LOCUS       sequence-299512-        6081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXK2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6081)
  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 6081)
  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-FOXK2
FEATURES             Location/Qualifiers
     source          1..6081
                     /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    2133..2267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2157..2176)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2211..2230
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2380..2400)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2380..2398)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2419..2435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2419..2435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2432..2454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2443..2459
                     /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(2467..2473,2474..2491,2492..2497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2512..2533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2650..2667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2821..3409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2901..2920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3580..4371,4372..4440))
                     /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     4203..4222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4441..4545)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4571..5026)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4708..4729)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4920..4939
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5097..6081
                     /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 tccgctgcag ccgcggcggc gttatctggc gcgggaacac caccagcagg aggaggagct
      181 ggaggaggag gagctggagg aggcggttct ccacctggtg gatgggctgt ggcacgcttg
      241 gagggccgcg agttcgagta tctgatgaag aagcgctcgg tgaccatcgg ccgcaactcg
      301 tcgcagggct cggtggacgt gagcatgggc cactcgagct tcatctcccg tcgtcacctc
      361 gaaatattta cacccccggg aggaggtggc catggcgggg ccgctccgga gctgccaccc
      421 gcgcagccaa gaccagatgc aggcggagat ttctacctgc gatgtttggg caagaacggg
      481 gtattcgtgg acggcgtgtt ccagaggcgc ggggcgccgc cgctgcagct gccgcgcgtg
      541 tgcacattca ggttcccgag cacaaacatc aagataacgt tcactgccct gtccagcgag
      601 aagagagaga agcaggaggc gtctgagtct ccagtgaagg ccgtacagcc acacatctcg
      661 cccctgacca tcaacattcc agacaccatg gcccacctca tcagccctct gccctccccc
      721 acgggaacca tcagcgctgc aaactcctgc ccctccagcc cccgtggagc ggggtcttca
      781 gggtacaagg tgggccgagt gatgccatct gacctcaatt taatggctga caactcacag
      841 cctgaaaatg aaaaggaagc ttcaggtgga gacagcccga aggatgattc aaagccgcct
      901 tactcctacg cgcagctgat agttcaggcg attacgatgg ctcccgacaa acagctcacc
      961 ctgaacggga tttatacaca catcactaaa aattatccct actacaggac tgcggacaag
     1021 ggctggcaga attcaattcg ccacaatctc tctctgaatc gttatttcat caaagtgccg
     1081 cgttcccagg aagaaccagg caaaggctcg ttctggagga tagacccagc ctctgaaagc
     1141 aaattaatag aacaggcttt taggaaacga cggcctaggg gcgtgccctg ctttagaacc
     1201 cctctgggac cgctctcttc taggagtgcc ccagcctctc ccaatcacgc gggagtgctg
     1261 tctgctcact ctagtggcgc ccagacccct gagagcctgt cgagggaagg ttcgccggcc
     1321 ccccttgagc ctgagcctgg cgctgcacag cccaaactcg ctgtcatcca ggaagcccgg
     1381 tttgcccaga gcgccccagg gtcacctctg tccagtcagc cagtcttaat caccgtccag
     1441 cggcagctac cacaggccat caagcctgtc acctacactg tggccacccc agtgaccacc
     1501 tcgacctccc agccacccgt cgtgcagacg gttcacgtcg tccaccagat cccagcggtg
     1561 tcggtcacca gtgtggccgg actggcccca gcgaacacgt acactgtctc tggacaagct
     1621 gtggtcaccc cggcagccgt gctggcccct cctaaggcag aggcccagga gaatggcgac
     1681 cacagggaag tcaaagtgaa agtagagcct attcccgcca ttggccacgc cacgctcggc
     1741 actgccagcc ggatcattca gacggcacag accaccccgg tccagacggt gaccatagta
     1801 caacaggcac ctctaggtca acaccagcta ccaataaaaa ctgtaacaca aaacggcact
     1861 cacgtggcat cagtccccac tgcggtccac ggccaggtga acaatgccgc ggcgagtcct
     1921 ttgcacatgt tggcaacaca cgcatccgca tcggcctccc tgcccacaaa gcgccacaac
     1981 ggtgaccagc cggagcagcc ggagctgaag cggatcaaga cagaagacgg cgagggcatc
     2041 gtcattgccc tgagcgtgga cacgccaccg gcagccgtaa gggaaaaggg tgtccagaac
     2101 tagtctagaa ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg
     2161 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2221 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2281 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg
     2341 cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     2401 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2461 tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2521 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2581 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2641 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2701 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3001 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3241 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4561 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4621 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4681 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4741 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4801 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4861 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4921 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4981 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     5041 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg
     5101 accgccaatt caatatggcg tatatggact catgccaatt caatatggtg gatctggacc
     5161 tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc
     5221 ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac
     5281 tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc
     5341 ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta
     5401 tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg
     5461 ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct
     5521 attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa
     5581 taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt
     5641 gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg
     5701 gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt
     5761 caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga
     5821 cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat
     5881 tactattgga agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa
     5941 atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg
     6001 caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata
     6061 agcaatgctc gtttagggaa c
//
