LOCUS       sequence-299001-        5802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXC1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5802)
  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 5802)
  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-FOXC1
FEATURES             Location/Qualifiers
     source          1..5802
                     /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    1854..1988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1878..1897)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1932..1951
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2101..2121)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2101..2119)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2140..2156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2140..2156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2153..2175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2164..2180
                     /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(2188..2194,2195..2212,2213..2218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2233..2254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2371..2388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2542..3130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2622..2641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3301..4092,4093..4161))
                     /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     3924..3943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4162..4266)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4292..4747)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4429..4450)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4641..4660
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4818..5802
                     /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 tcgaattcca ggcgcgctac tccgtgtcca gccccaactc cctgggagtg
      181 gtgccctacc tcggcggcga gcagagctac taccgcgcgg cggccgcggc ggccgggggc
      241 ggctacaccg ccatgccggc ccccatgagc gtgtactcgc accctgcgca cgccgagcag
      301 tacccgggcg gcatggcccg cgcctacggg ccctacacgc cgcagccgca gcccaaggac
      361 atggtgaagc cgccctatag ctacatcgcg ctcatcacca tggccatcca gaacgccccg
      421 gacaagaaga tcaccctgaa cggcatctac cagttcatca tggaccgctt ccccttctac
      481 cgggacaaca agcagggctg gcagaacagc atccgccaca acctctcgct caacgagtgc
      541 ttcgtcaagg tgccgcgcga cgacaagaag ccgggcaagg gcagctactg gacgctggac
      601 ccggactcct acaacatgtt cgagaacggc agcttcctgc ggcggcggcg gcgcttcaag
      661 aagaaggacg cggtgaagga caaggaggag aaggacaggc tgcacctcaa ggagccgccc
      721 ccgcccggcc gccagccccc gcccgcgccg ccggagcagg ccgacggcaa cgcgcccggt
      781 ccgcagccgc cgcccgtgcg catccaggac atcaagaccg agaacggtac gtgcccctcg
      841 ccgccccagc ccctgtcccc ggccgccgcc ctgggcagcg gcagcgccgc cgcggtgccc
      901 aagatcgaga gccccgacag cagcagcagc agcctgtcca gcgggagcag ccccccgggc
      961 agcctgccgt cggcgcggcc gctcagcctg gacggtgcgg attccgcgcc gccgccgccc
     1021 gcgccctccg ccccgccgcc gcaccatagc cagggcttca gcgtggacaa catcatgacg
     1081 tcgctgcggg ggtcgccgca gagcgcggcc gcggagctca gctccggcct tctggcctcg
     1141 gcggccgcgt cctcgcgcgc ggggatcgca cccccgctgg cgctcggcgc ctactcgccc
     1201 ggccagagct ccctctacag ctccccctgc agccagacct ccagcgcggg cagctcgggc
     1261 ggcggcggcg gcggcgcggg ggccgcgggg ggcgcgggcg gcgccgggac ctaccactgc
     1321 aacctgcaag ccatgagcct gtacgcggcc ggcgagcgcg ggggccactt gcagggcgcg
     1381 cccgggggcg cgggcggctc ggccgtggac gaccccctgc ccgactactc tctgcctccg
     1441 gtcaccagca gcagctcgtc gtccctgagt cacggcggcg gcggcggcgg cggcggggga
     1501 ggccaggagg ccggccacca ccctgcggcc caccaaggcc gcctcacctc gtggtacctg
     1561 aaccaggcgg gcggagacct gggccacttg gcgagcgcgg cggcggcggc ggcggccgca
     1621 ggctacccgg gccagcagca gaacttccac tcggtgcggg agatgttcga gtcacagagg
     1681 atcggcttga acaactctcc agtgaacggg aatagtagct gtcaaatggc cttcccttcc
     1741 agccagtctc tgtaccgcac gtccggagct ttcgtctacg actgtagcaa gttttgacac
     1801 accctcaaag ccgaacctcg agcctctaga actatagtga gtcgtattac gtagatccag
     1861 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1921 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1981 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2041 aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc
     2101 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2161 aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc
     2221 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2281 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2341 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2401 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2461 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2521 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2581 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2641 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2701 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2761 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2821 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2881 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2941 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3001 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3061 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3121 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3181 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3241 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3361 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3421 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3601 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4141 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4201 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4261 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     4321 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4381 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4441 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4501 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4561 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4621 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4681 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4741 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     4801 tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat
     4861 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
     4921 tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat
     4981 tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa
     5041 gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa
     5101 taatggcggc catattggct atatgccagg atcaatatat aggcaatatc caatatggcc
     5161 ctatgccaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
     5221 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
     5281 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
     5341 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
     5401 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
     5461 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
     5521 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
     5581 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
     5641 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
     5701 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
     5761 gcctaatggg aggtctatat aagcaatgct cgtttaggga ac
//
