LOCUS Exported 7486 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION GAL4-DNA binding domain fusion protein with the 329 N-terminal amino acids of human KLF4. ACCESSION . VERSION . KEYWORDS pCEFL GAL4dbd-KLF4 1-329 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7486) AUTHORS Yuan Y., Park J., Feng A., Awasthi P., Wang Z., Chen Q., Iglesias-Bartolome R. TITLE YAP1/TAZ-TEAD transcriptional networks maintain skin homeostasis by regulating cell proliferation and limiting KLF4 activity JOURNAL Nat Commun 11, 1472 (2020). REFERENCE 2 (bases 1 to 7486) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7486 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..212 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" intron 952..1155 /label=EF-1-alpha intron A /note="truncated intron upstream of the start codon of human EF-1-alpha" primer_bind 1112..1132 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1229..1669 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="DNA binding domain of the GAL4 transcriptional activator" /label=GAL4 DNA binding domain /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS" promoter complement(2723..2741) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2724..2741) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2761..2778) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2767..2991 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3037..3465 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3124..3143) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3334..3355 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3474..3494) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3479..3808 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3659..3794 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3721..3740 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3875..4669 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3929..3948) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4539..4558 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4843..4964 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4880..4899) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4934..4953 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5013..5029) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5013..5029) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5026..5048) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5037..5053 /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(5061..5091) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5106..5127 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5244..5261) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5415..6003) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5495..5514) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6174..7034) /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 6797..6816 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7035..7139) /gene="bla" /label=AmpR promoter primer_bind complement(7218..7237) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa 61 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc 121 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc 181 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc 241 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt 301 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg 361 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc 421 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca 481 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg 541 cgggccaaga tcgatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg 601 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga 661 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg 721 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa 781 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga 841 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct 901 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt 961 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact 1021 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt 1081 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt 1141 tttcttccat ttcaggtgtc gtgaggaatt ggcgaagctc agttaacggc cattgtggcc 1201 atcaagcttg aagcaagcct cctgaaagat gaagctactg tcttctatcg aacaagcatg 1261 cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg 1321 tctgaagaac aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag 1381 ggcacatctg acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat 1441 ttttcctcga gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc 1501 attgttaaca ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt 1561 ggcttcagtg gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc 1621 atcatcggaa gagagtagta acaaaggtca aagacagttg actgtatcgc cggaatttgg 1681 atccatgagg cagccacctg gcgagtctga catggctgtc agcgacgcgc tgctcccatc 1741 tttctccacg ttcgcgtctg gcccggcggg aagggagaag acactgcgtc aagcaggtgc 1801 cccgaataac cgctggcggg aggagctctc ccacatgaag cgacttcccc cagtgcttcc 1861 cggccgcccc tatgacctgg cggcggcgac cgtggccaca gacctggaga gcggcggagc 1921 cggtgcggct tgcggcggta gcaacctggc gcccctacct cggagagaga ccgaggagtt 1981 caacgatctc ctggacctgg actttattct ctccaattcg ctgacccatc ctccggagtc 2041 agtggccgcc accgtgtcct cgtcagcgtc agcctcctct tcgtcgtcgc cgtcgagcag 2101 cggccctgcc agcgcgccct ccacctgcag cttcacctat ccgatccggg ccgggaacga 2161 cccgggcgtg gcgccgggcg gcacgggcgg aggcctcctc tatggcaggg agtccgctcc 2221 ccctccgacg gctcccttca acctggcgga catcaacgac gtgagcccct cgggcggctt 2281 cgtggccgag ctcctgcggc cagaattgga cccggtgtac attccgccgc agcagccgca 2341 gccgccaggt ggcgggctga tgggcaagtt cgtgctgaag gcgtcgctga gcgcccctgg 2401 cagcgagtac ggcagcccgt cggtcatcag cgtcagcaaa ggcagccctg acggcagcca 2461 cccggtggtg gtggcgccct acaacggcgg gccgccgcgc acgtgcccca agatcaagca 2521 ggaggcggtc tcttcgtgca cccacttggg cgctggaccc cctctcagca atggccaccg 2581 gccggctgca cacgacttcc ccctggggcg gcagctcccc agcaggacta ccccgaccct 2641 gggtcttgag gaagtgctga gcagcaggga ctaagcggcc gcactcgaga tatcggcctc 2701 cagggcctct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag 2761 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 2821 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 2881 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 2941 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 3001 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 3061 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 3121 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 3181 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 3241 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 3301 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 3361 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 3421 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 3481 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 3541 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 3601 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 3661 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 3721 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 3781 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 3841 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 3901 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 3961 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 4021 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 4081 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 4141 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 4201 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 4261 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 4321 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 4381 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 4441 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 4501 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 4561 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 4621 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 4681 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 4741 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4801 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 4861 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 4921 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 4981 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 5041 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 5101 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 5161 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 5221 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 5281 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 5341 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 5401 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 5461 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 5521 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 5581 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 5641 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 5701 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 5761 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 5821 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 5881 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 5941 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 6001 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 6061 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 6121 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 6181 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 6241 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 6301 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 6361 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 6421 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 6481 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 6541 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 6601 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 6661 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 6721 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 6781 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 6841 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 6901 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 6961 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 7021 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 7081 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 7141 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcgatc tcccgatccc 7201 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc 7261 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg 7321 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt 7381 cgagctttgc aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc 7441 ttctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagt //