LOCUS       sequence-298999-        5420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXB2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5420)
  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 5420)
  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-FOXB2
FEATURES             Location/Qualifiers
     source          1..5420
                     /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             568..588
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             598..618
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    1472..1606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1496..1515)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1550..1569
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1719..1739)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1719..1737)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1758..1774)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1758..1774)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1771..1793)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1782..1798
                     /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(1806..1812,1813..1830,1831..1836))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1851..1872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1989..2006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2160..2748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2240..2259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2919..3710,3711..3779))
                     /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     3542..3561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3780..3884)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3910..4365)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4047..4068)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4259..4278
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4436..5420
                     /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 tcgaattccc gcggccgggg aagagctcgt acagcgacca aaaaccgccc
      181 tactcttaca tctcgctgac cgccatggca atccagcact cggccgagaa gatgctgccg
      241 ctgagcgaca tctacaagtt catcatggag cgcttcccct actaccgcga gcacacacag
      301 cgctggcaga acagcctgcg ccacaacctc tccttcaacg actgcttcat caagattccg
      361 cggaggcccg accagcctgg caagggtagc ttctgggcgc tgcaccccga ctgcggggac
      421 atgttcgaga acggcagctt cctgcggcgt cgcaagcgct tcaaggtgct gcgcgccgac
      481 catactcact tgcacgcggg aagcaccaag agcgcgccgg gcgccggtcc gggagggcac
      541 cttcaccccc atcaccacca ccacccccac caccaccatc atcaccacgc tgccgcacac
      601 caccaccatc accaccaccc accccagccg ccgccgccgc cgcccccgcc gccgccgcac
      661 atggtacact atttccatca gcaaccgcct actgctccgc agccgcctcc gcacctcccg
      721 tcacagcccc cgcagcaacc gccccagcag tcgcagcctc agcagccgtc tcaccccggc
      781 aagatgcagg aggcggcggc cgtggcggcg gcggcggcgg cggccgcggc agccgcggtg
      841 ggcagcgtgg gacgcctgtc tcagttccca ccctacgggc tgggctcggc cgccgccgct
      901 gccgccgcgg ccgcggcgtc cacgtcaggc ttcaagcacc cctttgccat tgagaacatt
      961 attggccggg actacaaggg cgtgctgcag gctggagggc tgcccttggc gtccgtcatg
     1021 caccacctgg gctaccccgt gcccggccag cttggcaacg tcgtcagctc cgtgtggccg
     1081 cacgttggcg tcatggattc ggtggccgcc gccgcggccg ccgcagccgc agccggagtc
     1141 cctgtaggcc cggagtatgg ggccttcggg gtcccggtca agtccctgtg ccactcggca
     1201 agccagagcc tgcctgccat gccggtgccc atcaagccca cgcctgcgct gccgcccgtg
     1261 tccgcgctgc agccggggct cactgtcccc gcggcttcgc agcagcctcc ggcgccatcc
     1321 accgtgtgct ccgcggccgc ggcctcgccc gttgcctctc tgctggagcc cacagcccct
     1381 acctcggccg aaagcaaggg cggctccttg cactcggtgc tagtgcactc ctagctcgag
     1441 cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat acattgatga
     1501 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1561 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1621 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc
     1681 ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     1741 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1801 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1861 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1921 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1981 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2041 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2341 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2581 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2641 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2821 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2881 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2941 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3001 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3061 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3121 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3181 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3241 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3301 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3361 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3421 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3481 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3541 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3601 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3661 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3721 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3781 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3841 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3901 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3961 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4021 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4081 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4141 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4201 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4261 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4321 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     4381 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga
     4441 ccgccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
     4501 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
     4561 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact
     4621 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
     4681 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
     4741 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
     4801 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
     4861 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
     4921 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
     4981 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
     5041 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
     5101 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
     5161 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
     5221 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
     5281 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
     5341 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
     5401 gcaatgctcg tttagggaac
//
