LOCUS       sequence-305127-        5606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXG1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5606)
  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 5606)
  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-FOXG1
FEATURES             Location/Qualifiers
     source          1..5606
                     /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"
     CDS             274..303
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             277..306
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    1658..1792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1682..1701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1736..1755
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1905..1925)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1905..1923)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1944..1960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1944..1960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1957..1979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1968..1984
                     /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(1992..1998,1999..2016,2017..2022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2037..2058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2175..2192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2346..2934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2426..2445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3105..3896,3897..3965))
                     /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     3728..3747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3966..4070)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4096..4551)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4233..4254)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4445..4464
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4622..5606
                     /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 tcgaattcct ggacatggga gataggaaag aggtgaaaat gatccccaag
      181 tcctcgttca gcatcaacag cctggtgccc gaggcggtcc agaacgacaa ccaccacgcg
      241 agccacggcc accacaacag ccaccacccc cagcaccacc accaccacca ccaccatcac
      301 caccacccgc cgccgcccgc cccgcaaccg ccgccgccgc cgcagcagca gcagccgccg
      361 ccgccgccgc ccccggcacg gcagcccccc cagacgcggg gcgccccggc cgccgacgac
      421 gacaagggcc cccagcagct gctgctcccg ccgccgccac cgccaccacc ggccgccgcc
      481 ctggacgggg ctaaagcgga cgggctgggc ggcaagggcg agccgggcgg cgggccgggg
      541 gagctggcgc ccgtcgggcc ggacgagaag gagaagggcg ccggcgccgg gggggaggag
      601 aagaaggggg cgggcgaggg cggcaaggac ggggaggggg gcaaggaggg cgagaagaag
      661 aacggcaagt acgagaagcc gccgttcagc tacaacgcgc tcatcatgat ggccatccgg
      721 cagagccccg agaagcggct cacgctcaac ggcatctacg agttcatcat gaagaacttc
      781 ccttactacc gcgagaacaa gcagggctgg cagaactcca tccgccacaa tctgtccctc
      841 aacaagtgct tcgtgaaggt gccgcgccac tacgacgacc cgggcaaggg caactactgg
      901 atgctggacc cgtcgagcga cgacgtgttc atcggcggca ccacgggcaa gctgcggcgc
      961 cgctccacca cctcgcgggc caagctggcc ttcaagcgcg gtgcgcgcct cacctccacc
     1021 ggcctcacct tcatggaccg cgccggctcc ctctactggc ccatgtcgcc cttcctgtcc
     1081 ctgcaccacc cccgcgccag cagcactttg agttacaacg gcaccacgtc ggcctacccc
     1141 agccacccca tgccctacag ctccgtgttg actcagaact cgctgggcaa caaccactcc
     1201 ttctccaccg ccaacggcct gagcgtggac cggctggtca acggggagat cccgtacgcc
     1261 acgcaccacc tcacggccgc cgcgctagcc gcctcggtgc cctgcggcct gtcggtgccc
     1321 tgctctggga cctactccct caacccctgc tccgtcaacc tgctcgcggg ccagaccagt
     1381 tactttttcc cccacgtccc gcacccgtca atgacttcgc agagcagcac gtccatgagc
     1441 gccagggccg cgtcctcctc cacgtcgccg caggccccct cgaccctgcc ctgtgagtct
     1501 ttaagaccct ctttgccaag ttttacgacg ggactgtctg ggggactgtc tgattatttc
     1561 acacatcaaa atcaggggtc ttcttccaac cctttaatac attaacatcc ctgggaccag
     1621 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     1681 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1741 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1801 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     1861 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     1921 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1981 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     2041 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2101 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2161 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2221 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2281 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2341 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2401 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2461 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2521 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2581 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2641 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2701 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2761 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2821 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2881 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2941 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3001 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3061 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3121 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3181 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3241 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3301 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3361 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3421 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3481 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3541 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3601 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3661 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3721 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3781 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3841 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3901 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3961 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4021 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4081 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4141 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4201 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4261 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4321 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4381 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4441 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4501 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     4561 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4621 agtcgaccgc caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct
     4681 ggacctgtgc caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct
     4741 ggaccccagc caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct
     4801 ggcactgtgc caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct
     4861 tggccctgtg ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt
     4921 ggctatatgc caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct
     4981 attggccagg ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt
     5041 ttcctattga cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt
     5101 cctaataccg cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc
     5161 ctattgacgt catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa
     5221 ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt
     5281 gacgtcaatg ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc
     5341 aatgacggta aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag
     5401 tacattacta ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc
     5461 ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa
     5521 tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct
     5581 atataagcaa tgctcgttta gggaac
//
