LOCUS       sequence-299343-        5268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXF1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5268)
  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 5268)
  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-FOXF1
FEATURES             Location/Qualifiers
     source          1..5268
                     /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    1320..1454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1344..1363)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1398..1417
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1567..1587)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1567..1585)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1606..1622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1606..1622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1619..1641)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1630..1646
                     /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(1654..1660,1661..1678,1679..1684))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1699..1720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1837..1854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2008..2596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2088..2107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2767..3558,3559..3627))
                     /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     3390..3409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3628..3732)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3758..4213)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3895..3916)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4107..4126
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4284..5268
                     /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 tcgaattctc ttcggcgccc gagaagcagc agccaccgca cggcggcggc
      181 ggcggcggcg gcgggggagg cggcgcggcc atggaccccg cgtcgtccgg cccgtccaag
      241 gccaagaaga ccaacgccgg catccggcgc ccggagaagc cgccctattc ctacatcgcg
      301 ctcatcgtca tggccatcca gagttcaccc accaagcgcc tgacgctgag cgagatctac
      361 cagttcctgc agagccgctt ccccttcttc cggggctcct accagggctg gaagaactcc
      421 gtgcgccaca acctctcgct caacgagtgc ttcatcaagc tacccaaggg ccttgggcgg
      481 cccggcaagg gccactactg gaccatcgac ccggccagcg agttcatgtt cgaggagggc
      541 tcctttcggc ggcggccgcg cggcttccga aggaaatgcc aggcgctcaa gcccatgtac
      601 agcatgatga acgggctcgg cttcaaccac ctcccggaca cctacggctt ccagggctcg
      661 gccggcggcc tctcgtgccc gcccaacagc ctggcgctgg agggcggcct gggcatgatg
      721 aacggccact tgccgggcaa cgtggacggc atggccctgc ccagccactc ggtgccccac
      781 ctgccttcca acggcggcca ctcgtacatg ggcggctgcg gcggcgcggc ggccggcgag
      841 tacccgcacc acgacagctc ggtgcccgcc tccccgctgc tgcccaccgg cgccggtggg
      901 gtcatggagc cgcacgccgt ctactcgggc tcggcggcgg cctggccgcc ctcggcgtcc
      961 gcggcgctca acagcggcgc ctcttatatc aagcagcagc ccctgtcccc ctgtaacccc
     1021 gcggccaacc ccctgtccgg cagcctctcc acgcactccc tggagcagcc gtatctgcac
     1081 cagaacagcc acaacgcccc agccgagctg caaggcatcc cgcggtatca ctcgcagtcg
     1141 cccagcatgt gtgaccgaaa ggagtttgtc ttctctttca acgccatggc gtcctcttcc
     1201 atgcactcgg ccggcggggg ctcctactac caccagcagg tcacctacca agacatcaag
     1261 ccttgcgtga tgtgaggctg ccctcgagcc tctagaacta tagtgagtcg tattacgtag
     1321 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1381 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1441 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1501 tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt
     1561 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     1621 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     1681 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1741 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1801 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1861 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1921 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1981 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2041 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2101 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2161 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2221 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2281 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2341 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2401 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2461 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2521 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2581 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2641 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2701 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2761 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2941 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3061 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3181 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3361 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3421 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3601 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3661 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3721 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     3781 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3841 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3901 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3961 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4021 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4081 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4141 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4201 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     4261 cgggcctctt cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat
     4321 gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt
     4381 gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat
     4441 gccaattcaa tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg
     4501 tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg
     4561 tgccaataat ggcggccata ttggctatat gccaggatca atatataggc aatatccaat
     4621 atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca
     4681 tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc
     4741 ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa
     4801 tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg
     4861 gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac
     4921 gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt
     4981 tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat
     5041 ctattaatag taacttggca agtacattac tattggaagt acgccagggt acattggcag
     5101 tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat
     5161 tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt
     5221 aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaac
//
