LOCUS       sequence-288278-        7012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL4-DNA binding domain fusion protein with amino acids 159 to 329 
            of human KLF4 (repressor domain).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7012)
  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. 2020 Mar 19;11(1):1472. doi: 10.1038/s41467-020-15301-0.
  PUBMED    32193376
REFERENCE   2  (bases 1 to 7012)
  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: "2020-03-19"; volume: "11(1):1472. doi"; pages: " 
            10.1038/s41467-020-15301-0"
COMMENT     Sequence Label: pCEFL GAL4dbd-KLF4 159-329
FEATURES             Location/Qualifiers
     source          1..7012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          213..1155
                     /label=EF-1-alpha intron A
                     /note="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(2249..2267)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2250..2267)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2287..2304)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2293..2517
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2563..2991
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2650..2669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2860..2881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3000..3020)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3005..3334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3185..3320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3247..3266
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3401..4195
                     /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(3455..3474)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4065..4084
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4369..4490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4406..4425)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4460..4479
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4539..4555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4539..4555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4552..4574)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4563..4579
                     /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(4587..4593,4594..4611,4612..4617))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4632..4653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4770..4787)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4941..5529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5021..5040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5700..6491,6492..6560))
                     /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     6323..6342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6561..6665)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6744..6763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6994..7012,1..1164)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
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 atccgacccg ggcgtggcgc cgggcggcac gggcggaggc ctcctctatg gcagggagtc
     1741 cgctccccct ccgacggctc ccttcaacct ggcggacatc aacgacgtga gcccctcggg
     1801 cggcttcgtg gccgagctcc tgcggccaga attggacccg gtgtacattc cgccgcagca
     1861 gccgcagccg ccaggtggcg ggctgatggg caagttcgtg ctgaaggcgt cgctgagcgc
     1921 ccctggcagc gagtacggca gcccgtcggt catcagcgtc agcaaaggca gccctgacgg
     1981 cagccacccg gtggtggtgg cgccctacaa cggcgggccg ccgcgcacgt gccccaagat
     2041 caagcaggag gcggtctctt cgtgcaccca cttgggcgct ggaccccctc tcagcaatgg
     2101 ccaccggccg gctgcacacg acttccccct ggggcggcag ctccccagca ggactacccc
     2161 gaccctgggt cttgaggaag tgctgagcag cagggactaa gcggccgcac tcgagatatc
     2221 ggcctccagg gcctctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     2281 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2341 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2401 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2461 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2521 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2581 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2641 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2701 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2761 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2821 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2881 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2941 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3001 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3061 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3181 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3241 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3301 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3361 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3421 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3481 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3541 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3601 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3661 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3721 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3781 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3841 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3901 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3961 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4021 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4081 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4141 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4201 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4261 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4321 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4381 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4441 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4501 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4561 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4621 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4681 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4741 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4801 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4861 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4921 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4981 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5041 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5101 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5161 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5221 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5281 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5341 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5401 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5461 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5521 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5581 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5641 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5701 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5761 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5821 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5881 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5941 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6001 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6061 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6121 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6181 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6241 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6301 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6361 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6421 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6481 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6541 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6601 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6661 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgatctccc
     6721 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6781 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     6841 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     6901 ctgcttcgag ctttgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa
     6961 tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca gt
//
