LOCUS       sequence-299513-        5249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXL2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5249)
  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 5249)
  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-FOXL2
FEATURES             Location/Qualifiers
     source          1..5249
                     /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    1301..1435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1325..1344)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1379..1398
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1548..1568)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1548..1566)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1587..1603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1587..1603)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1600..1622)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1611..1627
                     /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(1635..1641,1642..1659,1660..1665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1680..1701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1818..1835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1989..2577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2069..2088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2748..3539,3540..3608))
                     /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     3371..3390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3609..3713)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3739..4194)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3876..3897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4088..4107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4265..5249
                     /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 tcgaattcgc cagctacccc gagcccgagg acgcggcggg ggccctgctg
      181 gccccagaga ccggtcgcac agtcaaggag ccagaagggc cgccgccgag cccaggcaag
      241 ggcggtgggg gtggcggcgg gacagccccg gagaagccgg acccggcgca gaagcccccg
      301 tactcgtacg tggcgctcat cgccatggcg atccgcgaga gcgcggagaa gaggctcacg
      361 ctgtccggca tctaccagta catcatcgcg aagttcccgt tctacgagaa gaataagaag
      421 ggctggcaaa atagcatccg ccacaacctc agcctcaacg agtgcttcat caaggtgccg
      481 cgcgagggcg gcggcgagcg caagggcaac tactggacgc tggacccggc ctgcgaagac
      541 atgttcgaga agggcaacta ccggcgccgc cgccgcatga agaggccctt ccggccgccg
      601 cccgcgcact tccagcccgg caaggggctc ttcggggccg gaggcgccgc aggcgggtgc
      661 ggcgtggcgg gcgccggggc cgacggctac ggctacctgg cgccccccaa gtacctgcag
      721 tctggcttcc tcaacaactc gtggccgcta ccgcagcctc cctcacccat gccctatgcc
      781 tcctgccaga tggcggcagc cgcagcggct gcagcagctg cggctgcagc cgcgggcccc
      841 ggtagccctg gcgcggccgc tgtggtcaag gggctggcgg gcccggccgc ctcgtacggg
      901 ccgtacacac gcgtgcagag catggcgctg ccccccggcg tagtgaactc gtacaatggc
      961 ctgggaggcc cgccggccgc acccccgcct ccgccgcacc cccacccgca tccgcacgca
     1021 caccatctgc acgcggccgc cgcaccgccg cctgccccac cgcaccacgg ggccgccgcg
     1081 ccgccgccgg gccagctcag ccctgccagc ccagccaccg ccgcgccccc ggcgcccgcg
     1141 cccaccagtg cgccgggcct gcagttcgct tgtgcccggc agcccgagct cgccatgatg
     1201 cattgctctt actgggacca cgacagcaag accggcgcgc tgcattcgcg cctcgatctc
     1261 tgactcgagc ctctagaact atagtgagtc gtattacgta gatccagaca tgataagata
     1321 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1381 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1441 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt
     1501 cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg
     1561 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1621 ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa
     1681 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1741 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1801 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1861 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1921 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2401 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2641 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2701 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2761 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2821 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2881 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2941 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3001 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3061 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3121 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3181 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3241 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3301 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3361 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3421 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3481 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3541 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3601 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3661 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3721 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3781 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3841 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3901 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3961 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4021 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4081 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4141 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     4201 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4261 gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca atatggtgga
     4321 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga
     4381 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga
     4441 cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg
     4501 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat
     4561 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg
     4621 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg
     4681 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg
     4741 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct
     4801 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt
     4861 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc
     4921 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac
     4981 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc
     5041 aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat
     5101 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg
     5161 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg
     5221 tctatataag caatgctcgt ttagggaac
//
