LOCUS       sequence-370464-        7692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human KLF5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7692)
  AUTHORS   Goos H, Kinnunen M, Salokas K, Tan Z, Liu X, Yadav L, Zhang Q, Wei 
            GH, Varjosalo M
  TITLE     Human transcription factor protein interaction networks.
  JOURNAL   Nat Commun. 2022 Feb 9;13(1):766. doi: 10.1038/s41467-022-28341-5.
  PUBMED    35140242
REFERENCE   2  (bases 1 to 7692)
  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: "Nat 
            Commun."; date: "2022-02-9"; volume: "13(1):766. doi"; pages: " 
            10.1038/s41467-022-28341-5"
COMMENT     Sequence Label: MAC_N_KLF5
FEATURES             Location/Qualifiers
     source          1..7692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1999..2018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2190..2569
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2570..2773
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2724..2744
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2775..2793
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2796..2814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2824..2848
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3016..3366,3367..3369,3370..3978)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             3979..4005
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4036..4059
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4096..4119
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    4132..4156
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5537..5561)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5599..5616)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5605..5829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5962..5981)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6113..6160
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             6168..7190
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             6168..7190
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    7320..7441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7357..7376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7411..7430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7490..7506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7490..7506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7503..7525)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7514..7530
                     /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(7538..7544,7545..7562,7563..7568))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7583..7604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     1981 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     2041 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     2101 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     2161 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     2221 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2281 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2341 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2401 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2461 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2521 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2581 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2641 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2701 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2761 tataagcaga gctctcccta tcagtgatag agatctccct atcagtgata gagatcgtcg
     2821 acgagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     2881 ccatagaaga caccgggacc gatccagcct ccggactcta gcgtttaaac ttaagcttgg
     2941 taccgagctc ggatccacta gtccagtgtg gtggaattct gcagatatcc agcacagtgg
     3001 cggccgctcg agaccatgaa ggacaacacc gtgcccctga agctgatcgc cctgctggcc
     3061 aacggcgagt tccactctgg cgagcagctg ggcgagacac tgggcatgag cagagccgcc
     3121 atcaacaagc acatccagac cctgcgggac tggggcgtgg acgtgttcac agtgcccggc
     3181 aagggctaca gcctgcccga gcccatccag ctgctgaacg ccaagcagat cctgggccag
     3241 ctggatggcg gatctgtggc cgtgctgccc gtgatcgaca gcaccaacca gtatctgctc
     3301 gaccggatcg gcgagctgaa gtccggcgac gcctgtatcg ccgagtacca gcaggccggc
     3361 agaggcggca ggggcagaaa gtggttcagc cccttcggcg ccaacctgta cctgagcatg
     3421 ttctggagac tggaacaggg cccagccgcc gctattggcc tgagcctggt catcggcatc
     3481 gtgatggccg aggtgctgcg gaagctgggc gccgacaaag tgcgcgtgaa gtggcccaac
     3541 gacctgtatc tgcaggaccg gaagctggcc ggcatcctgg tggaactgac cggcaaaaca
     3601 ggcgacgccg cccagatcgt gatcggcgct ggcatcaaca tggccatgcg gcgggtggaa
     3661 gagagcgtgg tcaaccaggg ctggatcacc ctgcaggaag ccggcatcaa cctggaccgg
     3721 aacaccctgg ccgccatgct gatcagagag ctgagagccg ccctggaact gttcgagcag
     3781 gaaggcctgg ccccctacct gagcagatgg gagaagctgg acaacttcat caaccggccc
     3841 gtgaaactga tcatcggcga caaagagatc ttcggcatca gccggggcat cgacaagcag
     3901 ggggccctgc tcctggaaca ggacggcatc atcaagccct ggatgggcgg cgagatcagc
     3961 ctgcggagcg ccgagaagta cccatacgat gttcctgact atgccggtac cgagctcgga
     4021 tccaccatgg ctagctggag ccacccgcag ttcgagaaag gtggaggttc cggaggtgga
     4081 tcgggaggtg gatcgtggag ccacccgcag ttcgaaaaag cggccgatat cacaagtttg
     4141 tacaaaaaag caggctccac catggctaca agggtgctga gcatgagcgc ccgcctggga
     4201 cccgtgcccc agccgccggc gccgcaggac gagccggtgt tcgcgcagct caagccggtg
     4261 ctgggcgccg cgaatccggc ccgcgacgcg gcgctcttcc ccggcgagga gctgaagcac
     4321 gcgcaccacc gcccgcaggc gcagcccgcg cccgcgcagg ccccgcagcc ggcccagccg
     4381 cccgccaccg gcccgcggct gcctccagag gacctggtcc agacaagatg tgaaatggag
     4441 aagtatctga cacctcagct tcctccagtt cctataattc cagagcataa aaagtataga
     4501 cgagacagtg cctcagtcgt agaccagttc ttcactgaca ctgaagggtt accttacagt
     4561 atcaacatga acgtcttcct ccctgacatc actcacctga gaactggcct ctacaaatcc
     4621 cagagaccgt gcgtaacaca catcaagaca gaacctgttg ccattttcag ccaccagagt
     4681 gaaacgactg cccctcctcc ggccccgacc caggccctcc ctgagttcac cagtatattc
     4741 agctcacacc agaccgcagc tccagaggtg aacaatattt tcatcaaaca agaacttcct
     4801 acaccagatc ttcatctttc tgtccctacc cagcagggcc acctgtacca gctactgaat
     4861 acaccggatc tagatatgcc cagttctaca aatcagacag cagcaatgga cactcttaat
     4921 gtttctatgt cagctgccat ggcaggcctt aacacacaca cctctgctgt tccgcagact
     4981 gcagtgaaac aattccaggg catgccccct tgcacataca caatgccaag tcagtttctt
     5041 ccacaacagg ccacttactt tcccccgtca ccaccaagct cagagcctgg aagtccagat
     5101 agacaagcag agatgctcca gaatttaacc ccacctccat cctatgctgc tacaattgct
     5161 tctaaactgg caattcacaa tccaaattta cccaccaccc tgccagttaa ctcacaaaac
     5221 atccaacctg tcagatacaa tagaaggagt aaccccgatt tggagaaacg acgcatccac
     5281 tactgcgatt accctggttg cacaaaagtt tataccgagt cttctcattt aaaagctcac
     5341 ctgaggactc acactggtga aaagccatac aagtgtacct gggaaggctg cgactggagg
     5401 ttcgcgcgat cggatgagct gacccgccac taccggaagc acacaggcgc caagcccttc
     5461 cagtgcgggg tgtgcaaccg cagcttctcg cgctctgacc acctggccct gcatatgaag
     5521 aggcaccaga acttggaccc agctttcttg tacaaagtgg tgacgtaagc taggggcccg
     5581 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     5641 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     5701 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     5761 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     5821 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     5881 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5941 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6001 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6061 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc
     6121 tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac
     6181 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     6241 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     6301 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     6361 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     6421 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     6481 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     6541 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     6601 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     6661 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     6721 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     6781 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     6841 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     6901 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     6961 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     7021 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     7081 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     7141 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac
     7201 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     7261 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     7321 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7381 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7441 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     7501 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     7561 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     7621 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7681 cggggagagg cg
//
