LOCUS       sequence-299514-        6368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXM1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6368)
  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 6368)
  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-FOXM1
FEATURES             Location/Qualifiers
     source          1..6368
                     /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"
     RBS             1408..1419
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     polyA_signal    2420..2554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2444..2463)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2498..2517
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2667..2687)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2667..2685)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2706..2722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2706..2722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2719..2741)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2730..2746
                     /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(2754..2760,2761..2778,2779..2784))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2799..2820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2937..2954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3108..3696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3188..3207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3867..4658,4659..4727))
                     /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     4490..4509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4728..4832)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4858..5313)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4995..5016)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5207..5226
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5384..6368
                     /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 tcgaattcaa aactagcccc cgtcggccac tgattctcaa aagacggagg
      181 ctgccccttc ctgttcaaaa tgccccaagt gaaacatcag aggaggaacc taagagatcc
      241 cctgcccaac aggagtctaa tcaagcagag gcctccaagg aagtggcaga gtccaactct
      301 tgcaagtttc cagctgggat caagattatt aaccacccca ccatgcccaa cacgcaagta
      361 gtggccatcc ccaacaatgc taatattcac agcatcatca cagcactgac tgccaaggga
      421 aaagagagtg gcagtagtgg gcccaacaaa ttcatcctca tcagctgtgg gggagcccca
      481 actcagcctc caggactccg gcctcaaacc caaaccagct atgatgccaa aaggacagaa
      541 gtgaccctgg agaccttggg accaaaacct gcagctaggg atgtgaatct tcctagacca
      601 cctggagccc tttgcgagca gaaacgggag acctgtgcag atggtgaggc agcaggctgc
      661 actatcaaca atagcctatc caacatccag tggcttcgaa agatgagttc tgatggactg
      721 ggctcccgca gcatcaagca agagatggag gaaaaggaga attgtcacct ggagcagcga
      781 caggttaagg ttgaggagcc ttcgagacca tcagcgtcct ggcagaactc tgtgtctgag
      841 cggccaccct actcttacat ggccatgata caattcgcca tcaacagcac tgagaggaag
      901 cgcatgactt tgaaagacat ctatacgtgg attgaggacc actttcccta ctttaagcac
      961 attgccaagc caggctggaa gaactccatc cgccacaacc tttccctgca cgacatgttt
     1021 gtccgggaga cgtctgccaa tggcaaggtc tccttctgga ccattcaccc cagtgccaac
     1081 cgctacttga cattggacca ggtgtttaag cagcagaaac gaccgaatcc agagctccgc
     1141 cggaacatga ccatcaaaac cgaactcccc ctgggcgcac ggcggaagat gaagccactg
     1201 ctaccacggg tcagctcata cctggtacct atccagttcc cggtgaacca gtcactggtg
     1261 ttgcagccct cggtgaaggt gccattgccc ctggcggctt ccctcatgag ctcagagctt
     1321 gcccgccata gcaagcgagt ccgcattgcc cccaaggtgc tgctagctga ggaggggata
     1381 gctcctcttt cttctgcagg accagggaaa gaggagaaac tcctgtttgg agaagggttt
     1441 tctcctttgc ttccagttca gactatcaag gaggaagaaa tccagcctgg ggaggaaatg
     1501 ccacacttag cgagacccat caaagtggag agccctccct tggaagagtg gccctccccg
     1561 gccccatctt tcaaagagga atcatctcac tcctgggagg attcgtccca atctcccacc
     1621 ccaagaccca agaagtccta cagtgggctt aggtccccaa cccggtgtgt ctcggaaatg
     1681 cttgtgattc aacacaggga gaggagggag aggagccggt ctcggaggaa acagcatcta
     1741 ctgcctccct gtgtggatga gccggagctg ctcttctcag aggggcccag tacttcccgc
     1801 tgggccgcag agctcccgtt cccagcagac tcctctgacc ctgcctccca gctcagctac
     1861 tcccaggaag tgggaggacc ttttaagaca cccattaagg aaacgctgcc catctcctcc
     1921 accccgagca aatctgtcct ccccagaacc cctgaatcct ggaggctcac gcccccagcc
     1981 aaagtagggg gactggattt cagcccagta caaacccccc agggtgcctc tgaccccttg
     2041 cctgaccccc tggggctgat ggatctcagc accactccct tgcaaagtgc tccccccctt
     2101 gaatcaccgc aaaggctcct cagttcagaa cccttagacc tcatctccgt cccctttggc
     2161 aactcttctc cctcagatat agacgtcccc aagccaggct ccccggagcc acaggtttct
     2221 ggccttgcag ccaatcgttc tctgacagaa ggcctggtcc tggacacaat gaatgacagc
     2281 ctcagcaaga tcctgctgga catcagcttt cctggcctgg acgaggaccc actgggccct
     2341 gacaacatca actggtccca gtttattcct gagctacagt agctcgagcc tctagaacta
     2401 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
     2461 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2521 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2581 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
     2641 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     2701 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2761 cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2941 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3001 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3901 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3961 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4021 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4081 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4141 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4201 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4261 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4321 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4381 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4441 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4501 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4561 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4621 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4681 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4741 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4801 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     4861 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4921 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4981 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5041 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5101 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5161 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5221 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5281 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     5341 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc gccaattcaa
     5401 tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa
     5461 tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa
     5521 tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt gccaactggg
     5581 gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc
     5641 gccatattga attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca
     5701 atatataggc aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac
     5761 tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata
     5821 gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc
     5881 cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg
     5941 cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc
     6001 ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct
     6061 cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc
     6121 acttggcagt acatcaatat ctattaatag taacttggca agtacattac tattggaagt
     6181 acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg
     6241 gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc
     6301 cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt
     6361 tagggaac
//
