LOCUS       sequence-299511-        6317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXK1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6317)
  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 6317)
  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-FOXK1
FEATURES             Location/Qualifiers
     source          1..6317
                     /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    2369..2503
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2393..2412)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2447..2466
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2616..2636)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2616..2634)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2655..2671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2655..2671)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2668..2690)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2679..2695
                     /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(2703..2709,2710..2727,2728..2733))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2748..2769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2886..2903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3057..3645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3137..3156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3816..4607,4608..4676))
                     /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     4439..4458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4677..4781)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4807..5262)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4944..4965)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5156..5175
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5333..6317
                     /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 tcgctgaagt cggcgaggac agcggcgcaa gagccctgct tgcgctacgc
      181 tcggctccat gcagcccagt tctgtgtgca gcagcagctg cagctgcctt tcctgctgct
      241 gcaccacctc cagcacccgc tcagccacaa cctcctccag gtcctccacc accaccacca
      301 cctcctctac ctccaggagc tattgctggc gcgggctcat ctggtggttc ctctggggta
      361 tccggtgact cagcagtcgc aggagcggcg cctgcattag tggcagctgc agccgcatcg
      421 gtaaggcaga gtcctggtcc agcactggcg cgactagagg gacgcgagtt cgagttcctc
      481 atgcgccagc ctagcgtcac catcggccgc aactcgtcgc agggttcggt ggacttgagc
      541 atgggcctgt ccagcttcat ctcgcggcgc cacttgcagc tcagcttcca ggagccacac
      601 ttctacctgc gctgcctcgg taagaacggc gttttcgtgg acggtgcttt ccagagacgc
      661 ggcgcgcccg ccctgcagct gcccaagcag tgtaccttcc ggtttcccag cacggccatc
      721 aagatccagt tcacgtcgct ctatcacaaa gaagaggccc cagcctcccc gctgcggcca
      781 ctgtaccccc agatctcccc tctgaagatc cacatcccgg agccggacct tcggagtatg
      841 gttagccctg tcccttcccc gacgggcacc ataagtgtcc caaactcctg cccagccagt
      901 ccacgcggtg caggctcctc cagttaccgc tttgtgcaga acgtgacctc ggacctgcag
      961 ctggcagcag agtttgcagc aaaggccgcg tcggagcagc aggcagacac gtctggagga
     1021 gacagcccca aggatgagtc aaagccgccg ttctcctacg cgcagctgat cgtgcaggcc
     1081 atctcctccg cccaggaccg gcagctgacc ctgagcggga tctacgccca catcaccaag
     1141 cattacccct actaccggac ggccgacaaa ggctggcaga attctatccg gcacaacctc
     1201 tctttgaacc gttactttat caaagtccca cgttcccagg aggagcctgg gaaggggtcc
     1261 ttttggcgaa tagaccctgc ctctgaagcc aagctcgtgg aacaggcatt ccggaaacgg
     1321 aggcagaggg gtgtctcctg cttccgcacc cccttcgggc ctttgtcttc aaggagcgct
     1381 ccagcttcac ccacacaccc cggactgatg tcccctcgat ccggcggtct gcagacccca
     1441 gagtgtctgt ctcgggaggg atcccccatt ccacacgacc ctgagtttgg gtccaagtta
     1501 gcttctgtcc cagagtaccg gtattcccaa agcgcacctg gctctcccgt cagtgcccag
     1561 ccagtgatca tggccgtgcc tccccgaccg tccagtctcg tggctaagcc cgtagcctac
     1621 atgcctgcat ccatcgtaac ctcacagcag cccgctggcc acgccatcca cgtagtgcag
     1681 caggccccca ccgtcaccat ggtcagggta gtcaccacat ctgccaactc ggccaacgga
     1741 tacatcctca ctagccaggg tgcggcggga ggatcccatg atgcggcggg cgcagcagtt
     1801 ttggatttgg gcagcgaggc cagaggcctg gaggagaaac ccaccattgc gtttgccaca
     1861 atccctgcgg ctggtggagt catccagacg gtggccagcc agatggcacc cggggtaccc
     1921 ggacatacgg tcactatcct gcagcccgct acacccgtga ctctcgggca acaccatctt
     1981 ccagtccggg ccgtgaccca gaacggaaag catgcggttc ccacgaacag tttagccggc
     2041 aacgcttacg ccctcaccag ccctttgcag ctccttgcga cccaagcgag ttcatccgcg
     2101 ccggttgtgg tcactcgggt atgcgaagtg gggcccaagg aaccagcagc agctgtcgct
     2161 gcaacggcca ccactacccc agccactgct accacagcct ctgcatccgc ctcttccact
     2221 ggagagcccg aggtcaaaag gtcacgggta gaggagccca gtggggctgt aacaacaccg
     2281 gctggagtaa tcgcagctgc cggaccccag gggccaggaa ccggtgagta gctcgagcct
     2341 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     2401 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2461 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2521 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     2581 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     2641 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2701 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2761 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2821 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4741 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4801 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4861 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4921 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4981 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     5041 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5101 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     5161 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5221 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     5281 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg
     5341 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     5401 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     5461 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg
     5521 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     5581 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     5641 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     5701 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     5761 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     5821 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     5881 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     5941 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     6001 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     6061 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
     6121 attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg
     6181 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa
     6241 tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca
     6301 atgctcgttt agggaac
//
