LOCUS       sequence-299712-        5503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXD4L5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5503)
  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 5503)
  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-FOXD4L5
FEATURES             Location/Qualifiers
     source          1..5503
                     /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    1555..1689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1579..1598)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1633..1652
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1802..1822)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1802..1820)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1841..1857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1841..1857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1854..1876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1865..1881
                     /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(1889..1895,1896..1913,1914..1919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1934..1955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2072..2089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2243..2831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2323..2342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3002..3793,3794..3862))
                     /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     3625..3644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3863..3967)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3993..4448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4130..4151)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4342..4361
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4519..5503
                     /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 tcgaattcaa cttgccaaga gctgagcgcc ctcgctccac accgcagcgc
      181 agcctccggg actccgatgg ggaagacggt aaaatcgatg tcctgggaga ggaggaagat
      241 gaagacgagg tggaagacga ggaggaggag gcgagacagc agttcctaga gcagtcgctc
      301 cagccggggc tgcaggtggc ccggtggggc ggggttgcgc ttccccgaga gcacatcgag
      361 ggcggcggcg gcccgagtga cccttcagag tttggcacca agttcagggc accgccaagg
      421 tctgcggcgg cctctgaaga tgcccggcag ccggcaaagc ccccctactc gtacatcgcg
      481 ctcatcacca tggccatcct gcaaaacccg cacaagcgcc tcacgctcag cggcatctgc
      541 gccttcatta gtggccgctt cccatactac cgccgcaagt tccccgcctg gcagaacagc
      601 atccgccaca acctctcgct gaacgactgc ttcgttaaga tcccccgcga gccgggccac
      661 ccaggcaagg gcaactactg gagcctggac cccgcctccc aggacatgtt cgacaatggc
      721 agctttctcc ggcgtaggaa gcgtttcaag cgccaccaac tgaccccggg agcccacctg
      781 ccccacccct tccctctacc tgctgcacac gccgccctgc acaaccccca cccaggccct
      841 ctgcttgggg cccctgcccc gccgcagcca gtcccggggg cctaccccaa caccgccccc
      901 gggagacgcc cttacgctct gctgcacccg catcctcttc gctacctact gctctcggcc
      961 cccgtctatg ccggggcacc gaagaaagca gaaggcgcgg acctggcgac cccggcaccc
     1021 ttcccgtgct gcagccctca cttggtcctc agccttggga ggagggcaag ggtctggcgt
     1081 cgccaccggg aggcggatgc atctctttca gcattgagag tattatgcaa ggggtcaggg
     1141 gagcgggtac aggggctgcg cagagtttgt ccccgaccgc gtggagctac tgccacctgc
     1201 tccagcgacc atcaagcctg ttgcatcccc aaaccgctgc ccctttgctg caagtgtccg
     1261 ccgccgctgc tgctcggaca attttgcagc aatagcagca gcatcaggag gactgcgcca
     1321 acggctgcgc tcccaccaag ggcgcggtgc tgggcgggca cctgtcggcc tcgtcggcgc
     1381 tgaggtatca ggcagtggcg gagggctcta ggctgacatc gctggctgcc cctttgggcg
     1441 gagaggggac ctcaccagtt tttttagtat cgcccacgcc cagttccctg gccaactccg
     1501 cagggccctc ctagagcctc gagcctctag aactatagtg agtcgtatta cgtagatcca
     1561 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     1621 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     1681 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     1741 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     1801 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1861 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     1921 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1981 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2101 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2641 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3961 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     4021 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4081 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4141 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4201 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4261 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4321 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4381 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4441 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     4501 ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa
     4561 ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa
     4621 ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa
     4681 ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca
     4741 agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca
     4801 ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc
     4861 cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag
     4921 tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata
     4981 taccatatat ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc
     5041 tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc
     5101 tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa
     5161 taggaccacc caccattgac gtcaatggga tggctcattg cccattcata tccgttctca
     5221 cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt
     5281 aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc
     5341 ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg
     5401 tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg
     5461 tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
