LOCUS       sequence-299345-        5476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXF2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5476)
  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 5476)
  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-FOXF2
FEATURES             Location/Qualifiers
     source          1..5476
                     /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             931..951
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    1528..1662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1552..1571)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1606..1625
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1775..1795)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1775..1793)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1814..1830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1814..1830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1827..1849)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1838..1854
                     /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(1862..1868,1869..1886,1887..1892))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1907..1928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2045..2062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2216..2804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2296..2315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2975..3766,3767..3835))
                     /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     3598..3617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3836..3940)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3966..4421)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4103..4124)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4315..4334
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4492..5476
                     /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 tcgaattcac caccgagggc gggccgccgc cggccccgct ccgccgcgcg
      181 tgcagcccgg tccccggcgc gctccaggcc gccctgatga gcccgccgcc cgccgccgcc
      241 gccgccgccg ccgccgcccc ggagaccacc tcctcctcct cgtcgtcgtc ctccgcctcc
      301 tgcgcctcgt cctcgtcctc ctccaattcg gccagcgccc cctcggctgc ctgcaagagc
      361 gcgggcggcg gcggcgcggg cgccgggagc gggggcgcca agaaggcgag ctcggggctg
      421 cggcggcccg agaagccgcc ctactcgtac atcgcgctca tcgtcatggc catccagagc
      481 tcgcccagca agcgcctgac gctcagcgag atctaccagt tcctgcaggc gcgcttcccc
      541 ttcttccgcg gcgcctacca gggctggaag aactcggtgc gccacaatct ctcgctcaac
      601 gagtgcttca tcaagctgcc taagggcctc gggcggcccg gcaagggcca ctactggacc
      661 atcgacccgg ccagcgagtt catgttcgag gagggctcgt tccgccgccg gccgcgcggc
      721 ttcaggcgga agtgccaggc gctcaagccc atgtaccacc gcgtggtgag cggcttgggc
      781 ttcggggcgt cgctgctgcc ccagggcttc gacttccagg cgcccccgtc ggcgccgctc
      841 ggctgccaca gccagggcgg ctacggcggc ctcgacatga tgcccgcggg ctacgacgcc
      901 ggcgcgggcg cccccagcca cgcgcaccct caccaccacc accaccacca cgtcccgcac
      961 atgtcgccca acccgggttc cacctacatg gccagctgcc cggtgcccgc gggacccggg
     1021 ggcgtcggtg cggccggggg cggcggcggc ggcggcgact acgggccgga cagcagcagc
     1081 agcccggtac cctcgtcccc ggccatggcg agcgccatcg aatgccactc gccctacacg
     1141 agccctgcgg cgcactggag ctcgcctggc gcctcgcctt acctcaagca gccgcctgcc
     1201 ctgacgccca gcagcaaccc cgccgcctcg gcaggcctgc actccagcat gtcctcctac
     1261 tcgctggagc agagctactt gcaccagaac gctcgcgagg acctctcagt gggactgccc
     1321 cgttaccagc atcactctac tccagtgtgt gacagaaaag atttcgtcct caacttcaat
     1381 gggatttctt ctttccatcc ctcagctagc gggtcgtatt accaccatca ccaccagagc
     1441 gtctgtcagg atattaagcc ctgcgtcatg tgaacggaaa gaggccaagc ctcgagcctc
     1501 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
     1561 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1621 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1681 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg
     1741 ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     1801 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1861 acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1921 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1981 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2041 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2101 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2161 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2221 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2281 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2341 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2401 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2461 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2521 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2581 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2641 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2701 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2761 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2821 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2881 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2941 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3001 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3061 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3121 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3181 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3241 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3301 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3361 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3421 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3481 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3541 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3601 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3661 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3721 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3781 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3841 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3901 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3961 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4021 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4081 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4141 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4201 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4261 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4321 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4381 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     4441 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc
     4501 caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct ggacctgtgc
     4561 caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct ggaccccagc
     4621 caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct ggcactgtgc
     4681 caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct tggccctgtg
     4741 ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt ggctatatgc
     4801 caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct attggccagg
     4861 ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt ttcctattga
     4921 cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt cctaataccg
     4981 cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc ctattgacgt
     5041 catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa
     5101 tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt gacgtcaatg
     5161 ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc aatgacggta
     5221 aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag tacattacta
     5281 ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc
     5341 ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat
     5401 gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa
     5461 tgctcgttta gggaac
//
