LOCUS       sequence-299002-        5632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXC2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5632)
  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 5632)
  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-FOXC2
FEATURES             Location/Qualifiers
     source          1..5632
                     /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    1684..1818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1708..1727)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1762..1781
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1931..1951)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1931..1949)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1970..1986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1970..1986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1983..2005)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1994..2010
                     /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(2018..2024,2025..2042,2043..2048))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2063..2084
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2201..2218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2372..2960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2452..2471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3131..3922,3923..3991))
                     /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     3754..3773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3992..4096)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4122..4577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4259..4280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4471..4490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4648..5632
                     /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 tccgtgtccg accccaacgc cctgggagtg
      181 gtgccctacc tgagcgagca gaattactac cgggctgcgg gcagctacgg cggcatggcc
      241 agccccatgg gcgtctattc cggccacccg gagcagtaca gcgcggggat gggccgctcc
      301 tacgcgccct accaccacca ccagcccgcg gcgcctaagg acctggtgaa gccgccctac
      361 agctacatcg cgctcatcac catggccatc cagaacgcgc ccgagaagaa gatcaccttg
      421 aacggcatct accagttcat catggaccgc ttccccttct accgggagaa caagcagggc
      481 tggcagaaca gcatccgcca caacctctcg ctcaacgagt gcttcgtcaa ggtgccccgc
      541 gacgacaaga agcccggcaa gggcagttac tggaccctgg acccggactc ctacaacatg
      601 ttcgagaacg gcagcttcct gcggcgccgg cggcgcttca aaaagaagga cgtgtccaag
      661 gagaaggagg agcgggccca cctcaaggag ccgcccccgg cggcgtccaa gggcgccccg
      721 gccacccccc acctagcgga cgcccccaag gaggccgaga agaaggtggt gatcaagagc
      781 gaggcggcgt ccccggcgct gccggtcatc accaaggtgg agacgctgag ccccgagagc
      841 gcgctgcagg gcagcccgcg cagcgcggcc tccacgcccg ccggctcccc cgacggctcg
      901 ctgccggagc accacgccgc ggcgcccaac gggctgcctg gcttcagcgt ggagaacatc
      961 atgaccctgc gaacgtcgcc gccgggcgga gagctgagcc cgggggccgg acgcgcgggc
     1021 ctggtggtgc cgccgctggc gctgccctac gccgccgcgc cgcccgccgc ctacggccag
     1081 ccgtgcgctc agggcctgga ggccggggcc gccgggggct accagtgcag catgcgagcg
     1141 atgagcctgt acaccggggc cgagcggccg gcgcacatgt gcgtcccgcc cgccctggac
     1201 gaggccctct cggaccaccc gagcggcccc acgtcgcccc tgagcgctct caacctcgcc
     1261 gccggccagg agggcgcgct cgccgccacg ggccaccacc accagcacca cggccaccac
     1321 cacccgcagg cgccgccgcc cccgccggct ccccagcccc agccgacgcc gcagcccggg
     1381 gccgccgcgg cgcaggcggc ctcctggtat ctcaaccaca gcggggacct gaaccacctc
     1441 cccggccaca cgttcgcggc ccagcagcaa actttcccca acgtgcggga gatgttcaac
     1501 tcccaccggc tggggattga gaactcgacc ctcggggagt cccaggtgag tggcaatgcc
     1561 agctgccagc tgccctacag atccacgccg cctctctatc gccacgcagc cccctactcc
     1621 tacgactgca cgaaatactg acgtctctcg agcctctaga actatagtga gtcgtattac
     1681 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1741 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1801 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1861 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
     1921 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     1981 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa
     2041 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2101 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2161 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2221 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2281 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2341 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2401 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2461 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2521 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2581 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2641 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2701 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2761 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2821 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2881 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2941 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3001 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3061 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3121 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3181 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3241 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3301 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3361 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3421 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3481 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3541 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3601 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3661 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3721 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3781 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3841 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3901 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3961 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4021 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4081 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4141 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4201 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4261 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4321 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4381 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4441 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4501 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4561 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4621 ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac
     4681 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac
     4741 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca
     4801 ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc
     4861 acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc
     4921 atggtgccaa taatggcggc catattggct atatgccagg atcaatatat aggcaatatc
     4981 caatatggcc ctatgccaat atggctattg gccaggttca atactatgta ttggccctat
     5041 gccatatagt attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc
     5101 ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc cccattgacg
     5161 tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta
     5221 tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat
     5281 tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat
     5341 ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca
     5401 atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg
     5461 gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc
     5521 ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg
     5581 cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga ac
//
