LOCUS       sequence-370381-        7719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human KLF10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7719)
  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 7719)
  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_KLF10
FEATURES             Location/Qualifiers
     source          1..7719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..81)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(235..823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(315..334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(994..1785,1786..1854))
                     /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     1617..1636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1855..1959)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2034..2053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2225..2604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2605..2808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2759..2779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2810..2828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2831..2849
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2859..2883
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3051..3401,3402..3404,3405..4013)
                     /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             4014..4040
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4071..4094
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4131..4154
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    4167..4191
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5599..5623)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5661..5678)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5667..5891
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6024..6043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6175..6222
                     /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             6230..7252
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             6230..7252
                     /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    7382..7503
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7419..7438)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7473..7492
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7552..7568)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7552..7568)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7565..7587)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7576..7592
                     /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(7600..7606,7607..7624,7625..7630))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7645..7666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
       61 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      181 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      241 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      301 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      361 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      421 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      481 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      541 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      601 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      661 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      721 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      901 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      961 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     1141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1201 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1261 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1861 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1921 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1981 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     2041 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     2101 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     2161 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     2221 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     2281 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     2341 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2401 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2461 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2521 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2581 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2641 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2701 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2761 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt
     2821 gatagagatc tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag
     2881 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     2941 agcctccgga ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca
     3001 gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagacc atgaaggaca
     3061 acaccgtgcc cctgaagctg atcgccctgc tggccaacgg cgagttccac tctggcgagc
     3121 agctgggcga gacactgggc atgagcagag ccgccatcaa caagcacatc cagaccctgc
     3181 gggactgggg cgtggacgtg ttcacagtgc ccggcaaggg ctacagcctg cccgagccca
     3241 tccagctgct gaacgccaag cagatcctgg gccagctgga tggcggatct gtggccgtgc
     3301 tgcccgtgat cgacagcacc aaccagtatc tgctcgaccg gatcggcgag ctgaagtccg
     3361 gcgacgcctg tatcgccgag taccagcagg ccggcagagg cggcaggggc agaaagtggt
     3421 tcagcccctt cggcgccaac ctgtacctga gcatgttctg gagactggaa cagggcccag
     3481 ccgccgctat tggcctgagc ctggtcatcg gcatcgtgat ggccgaggtg ctgcggaagc
     3541 tgggcgccga caaagtgcgc gtgaagtggc ccaacgacct gtatctgcag gaccggaagc
     3601 tggccggcat cctggtggaa ctgaccggca aaacaggcga cgccgcccag atcgtgatcg
     3661 gcgctggcat caacatggcc atgcggcggg tggaagagag cgtggtcaac cagggctgga
     3721 tcaccctgca ggaagccggc atcaacctgg accggaacac cctggccgcc atgctgatca
     3781 gagagctgag agccgccctg gaactgttcg agcaggaagg cctggccccc tacctgagca
     3841 gatgggagaa gctggacaac ttcatcaacc ggcccgtgaa actgatcatc ggcgacaaag
     3901 agatcttcgg catcagccgg ggcatcgaca agcagggggc cctgctcctg gaacaggacg
     3961 gcatcatcaa gccctggatg ggcggcgaga tcagcctgcg gagcgccgag aagtacccat
     4021 acgatgttcc tgactatgcc ggtaccgagc tcggatccac catggctagc tggagccacc
     4081 cgcagttcga gaaaggtgga ggttccggag gtggatcggg aggtggatcg tggagccacc
     4141 cgcagttcga aaaagcggcc gatatcacaa gtttgtacaa aaaagttgcc atggaggaaa
     4201 gaatggaaat gatttctgaa aggccaaaag agagtatgta ttcctggaac aaaactgcag
     4261 agaaaagtga ttttgaagct gtagaagcac ttatgtcaat gagctgcagt tggaagtctg
     4321 attttaagaa atacgttgaa aacagacctg ttacaccagt atctgatttg tcagaggaag
     4381 agaatctgct tccgggaaca cctgattttc atacaatccc agcattttgt ttgactccac
     4441 cttacagtcc ttctgacttt gaaccctctc aagtgtcaaa tctgatggca ccagcgccat
     4501 ctactgtaca cttcaagtca ctctcagata ctgccaaacc tcacattgcc gcacctttca
     4561 aagaggaaga aaagagccca gtatctgccc ccaaactccc caaagctcag gcaacaagtg
     4621 tgattcgtca tacagctgat gcccagctat gtaaccacca gacctgccca atgaaagcag
     4681 ccagcatcct caactatcag aacaattctt ttagaagaag aacccaccta aatgttgagg
     4741 ctgcaagaaa gaacatacca tgtgccgctg tgtcaccaaa cagatccaaa tgtgagagaa
     4801 acacagtggc agatgttgat gagaaagcaa gtgctgcact ttatgacttt tctgtgcctt
     4861 cctcagagac ggtcatctgc aggtctcagc cagcccctgt gtccccacaa cagaagtcag
     4921 tgttggtctc tccacctgca gtatctgcag ggggagtgcc acctatgccg gtcatctgcc
     4981 agatggttcc ccttcctgcc aacaaccctg ttgtgacaac agtcgttccc agcactcctc
     5041 ccagccagcc accagccgtt tgcccccctg ttgtgttcat gggcacacaa gtccccaaag
     5101 gcgctgtcat gtttgtggta ccccagcccg ttgtgcagag ttcaaagcct ccggtggtga
     5161 gcccgaatgg caccagactc tctcccattg cccctgctcc tgggttttcc ccttcagcag
     5221 caaaagtcac tcctcagatt gattcatcaa ggataaggag tcacatctgt agccacccag
     5281 gatgtggcaa gacatacttt aaaagttccc atctgaaggc ccacacgagg acgcacacag
     5341 gagaaaagcc tttcagctgt agctggaaag gttgtgaaag gaggtttgcc cgttctgatg
     5401 aactgtccag acacaggcga acccacacgg gtgagaagaa atttgcgtgc cccatgtgtg
     5461 accggcggtt catgaggagt gaccatttga ccaagcatgc ccggcgccat ctatcagcca
     5521 agaagctacc aaactggcag atggaagtga gcaagctaaa tgacattgct ctacctccaa
     5581 cccctgctcc cacacagtgc ccaactttct tgtacaaagt ggtgacgtaa gctaggggcc
     5641 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     5701 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     5761 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     5821 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     5881 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     5941 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     6001 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     6061 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     6121 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt
     6181 cctattccga agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga
     6241 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     6301 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     6361 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     6421 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga
     6481 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     6541 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     6601 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     6661 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     6721 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     6781 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     6841 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     6901 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     6961 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     7021 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     7081 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     7141 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     7201 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact
     7261 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7321 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7381 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7501 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7561 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7621 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7681 actgcccgct ttccagtcgg gaaacctgtc gtgccagct
//
