LOCUS       sequence-171335-        7228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gal4-PRDM14 funsion plasmid for luciferase recruitment assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7228)
  AUTHORS   Tu S, Narendra V, Yamaji M, Vidal SE, Rojas LA, Wang X, Kim SY, 
            Garcia BA, Tuschl T, Stadtfeld M, Reinberg D
  TITLE     Co-repressor CBFA2T2 regulates pluripotency and germline 
            development.
  JOURNAL   Nature. 2016 Jun 8;534(7607):387-90. doi: 10.1038/nature18004.
  PUBMED    27281218
REFERENCE   2  (bases 1 to 7228)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2016-06-8- 8"; volume: "534"; issue: "7607"; pages:
            "387-90"
COMMENT     Sequence Label: hP14Gal4
FEATURES             Location/Qualifiers
     source          1..7228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        93..140
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             159..533
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    663..784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(700..719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     754..773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(833..849)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(833..849)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(846..868)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    857..873
                     /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(881..887,888..905,906..911))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    926..947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1064..1081)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1235..1823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1315..1334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1994..2785,2786..2854))
                     /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     2617..2636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2855..2959)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3034..3053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3225..3604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3605..3808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3759..3779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    3810..3828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3831..3849
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3859..3883
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3993..4433
                     /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"
     CDS             6165..6191
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6226..6243)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6232..6456
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6502..6930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6589..6608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6799..6820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6939..6959)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6944..7228,1..45)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7124..7228,1..31)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7186..7205
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
       61 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
      121 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg
      181 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
      241 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
      301 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
      361 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
      421 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
      481 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
      541 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
      601 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
      661 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      721 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      781 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
      841 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      901 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      961 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     1021 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1081 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1141 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1201 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1261 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1321 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1381 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1441 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1501 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1561 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1621 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1681 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1741 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1801 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1861 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1921 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1981 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2041 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2101 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2161 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2221 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2281 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2341 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2401 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2461 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2521 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2581 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2641 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2701 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2761 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2821 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2881 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2941 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     3001 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3061 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     3121 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     3181 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     3241 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     3301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     3361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     3421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     3481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     3541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     3601 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     3661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     3721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     3781 acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc tccctatcag
     3841 tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
     3901 acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga ctctagcgtt
     3961 taaacttaag cttgaagcaa gcctcctgaa agatgaagct actgtcttct atcgaacaag
     4021 catgcgatat ttgccgactt aaaaagctca agtgctccaa agaaaaaccg aagtgcgcca
     4081 agtgtctgaa gaacaactgg gagtgtcgct actctcccaa aaccaaaagg tctccgctga
     4141 ctagggcaca tctgacagaa gtggaatcaa ggctagaaag actggaacag ctatttctac
     4201 tgatttttcc tcgagaagac cttgacatga ttttgaaaat ggattcttta caggatataa
     4261 aagcattgtt aacaggatta tttgtacaag ataatgtgaa taaagatgcc gtcacagata
     4321 gattggcttc agtggagact gatatgcctc taacattgag acagcataga ataagtgcga
     4381 catcatcatc ggaagagagt agtaacaaag gtcaaagaca gttgactgta tcgccggaat
     4441 tcatggctct accccggcca agtgaggccg tgcctcagga caaggtgtgc tacccgccgg
     4501 agagcagccc gcagaacctg gccgcgtact acacgccttt cccgtcctat ggacactaca
     4561 gaaacagcct ggccaccgtg gaggaagact tccaaccttt ccggcagctg gaggccgcag
     4621 cgtctgctgc ccccgccatg ccccccttcc ccttccggat ggcgcctccc ttgctgagcc
     4681 cgggtctggg cctacagagg gagcctctct acgatctgcc ctggtacagc aagctgccac
     4741 cgtggtaccc aattccccac gtccccaggg aagtgccgcc cttcctgagc agcagccacg
     4801 agtacgcggg tgccagcagt gaagatctgg gccaccaaat cattggtggc gacaacgaga
     4861 gtggcccgtg ttgtggacct gacactttaa ttccaccgcc ccctgcggat gcttctctgt
     4921 tacctgaggg gctgaggacc tcccagttat taccttgctc acccagcaag cagtcagagg
     4981 atggtcccaa accctccaac caagaaggga agtcccctgc tcggttccag ttcacggagg
     5041 aggacctgca cttcgttctg tacggggtca ctcccagcct ggagcaccca gccagcctgc
     5101 accatgcgat ttcaggcctc ctggtccccc cagacagctc tggatctgat tctcttcctc
     5161 aaactctgga taaagactcc cttcaacttc cagaaggtct atgcctcatg cagacggtgt
     5221 ttggtgaagt cccacatttt ggtgtgttct gcagtagttt tatcgccaaa ggagtcaggt
     5281 ttgggccctt tcaaggtaaa gtggtcaatg ccagtgaagt gaagacctac ggagacaatt
     5341 ctgtgatgtg ggagatcttt gaagatggtc atttgagcca ctttatagat ggaaaaggag
     5401 gtacggggaa ctggatgtcc tatgtcaact gtgcccgctt ccccaaggag cagaacctag
     5461 ttgctgtgca gtgtcaaggg catatatttt atgagagctg caaagagatt catcagaacc
     5521 aagagctcct tgtgtggtat ggagactgct atgagaaatt tctggatatt cctgtgagcc
     5581 ttcaggtcac agagccgggg aagcagccat ctgggccctc tgaagagtct gcagaaggct
     5641 acagatgtga aagatgtggg aaggtattta cctacaaata ttacagagat aagcacctca
     5701 agtacacccc ctgtgtggac aagggcgata ggaaatttcc ctgttctctc tgcaaacgat
     5761 cctttgagaa gcgggaccgg cttcggatcc acattcttca tgttcatgag aagcaccggc
     5821 ctcacaagtg ttctacatgt gggaaatgtt tctctcagtc ttccagccta aacaaacaca
     5881 tgcgagtcca ctctggagac agaccatacc agtgtgtgta ttgtactaag aggttcacag
     5941 cctccagcat actccgcaca cacatcaggc agcactccgg ggagaagccc ttcaaatgca
     6001 agtactgtgg taaatctttt gcatcccatg ctgcccatga cagccatgtc cggcgttcac
     6061 acaaggagga cgatggctgc tcatgcagca tctgtgggaa aatcttctca gatcaagaaa
     6121 cattctactc ccacatgaag tttcatgaag actacgaatt catgtaccca tacgatgttc
     6181 cagattacgc ttgatctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     6241 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     6301 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     6361 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6421 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     6481 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6541 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6601 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6661 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6721 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6781 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6841 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6901 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     6961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     7021 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     7081 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     7141 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     7201 cgaggccgcc tctgcctctg agctattc
//
