LOCUS       sequence-305131-        5831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXJ2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5831)
  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 5831)
  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-FOXJ2
FEATURES             Location/Qualifiers
     source          1..5831
                     /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             complement(808..819)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1883..2017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1907..1926)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1961..1980
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2130..2150)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2130..2148)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2169..2185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2169..2185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2182..2204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2193..2209
                     /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(2217..2223,2224..2241,2242..2247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2262..2283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2400..2417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2571..3159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2651..2670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3330..4121,4122..4190))
                     /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     3953..3972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4191..4295)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4321..4776)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4458..4479)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4670..4689
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4847..5831
                     /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 tccgcttctg acctagagag tagcctcacc tccatagact ggctccccca gctgaccctc
      181 cgagctacca ttgagaagct tggaagtgcc tcccaggctg ggcctcccgg gagcagccgc
      241 aagtgttcac cagggtcacc cacagatcct aatgccaccc tgagcaaaga cgaggcagca
      301 gtgcaccagg acggcaagcc acgatacagc tatgccactc tcatcaccta tgccatcaac
      361 tcctctccag ccaagaagat gaccctcagc gagatttacc gctggatctg tgataacttc
      421 ccctattaca agaatgctgg cattggttgg aagaattcaa tacggcacaa cctttctctc
      481 aacaagtgtt tccggaaggt gcccagacct cgggatgacc ctgggaaggg ttcctattgg
      541 acaattgaca cctgccctga catttcccga aagagaagac accctccaga tgatgatctg
      601 tcccaagact caccagaaca ggaggcaagc aagagcccac ggggaggcgt tgcagggagt
      661 ggagaagcct cactgcctcc tgaggggaat ccgcagatgt cacttcagag ccccacatct
      721 atagccagct acagccaggg cacaggatct gtggatggtg gagcagtggc agcaggggct
      781 tcaggccgag aaagtgctga gggtccccct cccctctata acaccaacca tgactttaaa
      841 ttctcctact cagagatcaa ctttcaggat ctaagctggt ccttccgcaa cctctataag
      901 tccatgctgg agaagtcctc ttcctcctct cagcacggct tttcttctct cctgggggac
      961 atcccaccct cgaacaacta ctacatgtat cagcagcagc agccaccgcc acctcaacag
     1021 cagcagcagc agcagcagcc gccacagcca cctccccagc agtcccagcc acagcagcag
     1081 caggcacctg cccagggccc ctcagctgta gggggtgctc ctccactgca caccccaagc
     1141 acagatggtt gtaccccacc agggggaaag caagctgggg cggaaggcta tgggcctccc
     1201 cctgtaatgg ccatgcatcc acccccgctg cagcatggag gctaccaccc tcatcagcac
     1261 catccccact cccaccctgc ccagcagcca ccacctccac agccacaggc acaaggccag
     1321 gctcccatca acaacactgg ctttgccttt ccttctgact ggtgctctaa tattgactct
     1381 ttaaaggaaa gcttcaagat ggtgaatcgg ctcaattggt ccagcattga gcagtcacaa
     1441 ttctcagaac tgatggagag tctacgacag gcagagcaga agaactggac cctcgaccag
     1501 catcacattg ccaatctgtg tgactccctc aaccacttcc ttactcagac tggtcacgtg
     1561 ccccctcaag ggggtaccca ccgcccacca gcccctgccc gtattgctga ctcctgtgcc
     1621 ctcaccagtg gcaaacagga gtcagccatg agccaagtga actcttatgg gcacccacaa
     1681 gctccccacc tctaccctgg cccatcacca atgtacccaa tccccaccca ggactcagca
     1741 ggatacaatc gcccagcaca ccatatggtc cctcggccat cagtgccacc tcctggtgcc
     1801 aatgaggaga tccctgatga cttcgactgg gacttgatca cttagctcga gcctctagaa
     1861 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     1921 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1981 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2041 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
     2101 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     2161 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2221 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2281 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2341 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2401 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2461 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2761 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3001 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3181 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3241 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4201 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4261 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4321 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4381 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4441 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4501 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4561 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4621 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4681 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4741 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     4801 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accgccaatt
     4861 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
     4921 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
     4981 caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact
     5041 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
     5101 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
     5161 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
     5221 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
     5281 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
     5341 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
     5401 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
     5461 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
     5521 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
     5581 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
     5641 agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
     5701 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
     5761 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     5821 gtttagggaa c
//
