LOCUS       sequence-299335-        5243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXE1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5243)
  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 5243)
  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-FOXE1
FEATURES             Location/Qualifiers
     source          1..5243
                     /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    1295..1429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1319..1338)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1373..1392
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1542..1562)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1542..1560)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1581..1597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1581..1597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1594..1616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1605..1621
                     /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(1629..1635,1636..1653,1654..1659))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1674..1695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1812..1829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1983..2571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2063..2082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2742..3533,3534..3602))
                     /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     3365..3384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3603..3707)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3733..4188)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3870..3891)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4082..4101
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4259..5243
                     /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 tgccgagagc gggccgccgc cgccgcagcc ggaggtgctg
      181 gctaccgtga aggaagagcg cggcgagacg gcagcagggg ccggggtccc aggggaggcc
      241 acgggccgcg gggcgggcgg gcggcgccgc aagcgccccc tgcagcgcgg gaagccgccc
      301 tacagctaca tcgcgctcat cgccatggcc atcgcgcacg cgcccgagcg ccgcctcacg
      361 ctgggcggca tctacaagtt catcaccgag cgcttcccct tctaccgcga caaccccaaa
      421 aagtggcaga acagcatccg ccacaacctc acactcaacg actgcttcct caagatcccg
      481 cgcgaggccg gccgcccggg taagggcaac tactgggcgc ttgaccccaa cgcggaggac
      541 atgttcgaga gcggcagctt cctgcgccgc cgcaagcgct tcaagcgctc ggacctctcc
      601 acctacccgg cttacatgca cgacgcggcg gctgccgcag ccgccgccgc cgccgccgcc
      661 gccgccgccg ccatcttccc aggcgcggtg cccgccgcgc gcccccccta cccgggcgcc
      721 gtctatgcag gctacgcgcc gccgtcgctg gccgcgccgc ctccagtcta ctaccccgcg
      781 gcgtcgcccg gcccttgccg cgtcttcggc ctggttcctg agcggccgct cagcccagag
      841 ctggggcccg caccgtcggg gcccggcggc tcttgcgcct ttgcctccgc cggcgccccc
      901 gctaccacca ccggctacca gcccgcaggc tgcaccgggg cccggccggc caacccctcc
      961 gcctatgcgg ctgcctacgc gggccccgac ggcgcgtacc cgcagggcgc cggcagtgcg
     1021 atctttgccg ctgctggccg cctggcggga cccgcttcgc ccccagcggg cggcagcagt
     1081 ggcggcgtgg agaccacggt ggacttctac gggcgcacgt cgcccggcca gttcggagcg
     1141 ctgggagcct gctacaaccc tggcgggcag ctcggagggg ccagtgcagg cgcctaccat
     1201 gctcgccatg ctgccgctta tcccggtggg atagatcggt tcgtgtccgc catgtgactc
     1261 gagcctctag aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga
     1321 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1381 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1441 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc
     1501 cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt
     1561 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1621 attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg
     1681 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1741 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1801 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1861 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1921 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1981 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2041 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2101 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2161 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2221 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2281 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2341 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2401 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2461 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2521 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2581 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2641 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2701 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2761 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2821 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2881 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2941 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3001 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3061 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3121 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3181 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3241 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3301 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3361 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3421 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3481 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3541 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3601 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3661 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3721 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3781 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3841 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3901 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3961 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4021 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4081 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4141 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     4201 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt
     4261 cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga
     4321 cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga
     4381 ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc
     4441 actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg
     4501 ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc
     4561 tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt
     4621 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
     4681 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
     4741 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
     4801 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
     4861 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
     4921 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
     4981 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
     5041 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
     5101 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
     5161 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
     5221 taagcaatgc tcgtttaggg aac
//
