LOCUS       sequence-485636-        8276 bp    DNA     circular SYN 09-SEP-2025
DEFINITION  Expression of full-length human SF-1 using piggyBac transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8276)
  AUTHORS   Chan YS, Gao Q, Robinson SA, Wang W, Filandrova R, Weinhold LM, 
            Cabrera ML, Zhang M, Ambati CSR, Lerario AM, Putluri N, 
            Kiseljak-Vassiliades K, Wierman ME, Habra MA, Hammer GD, Veverka V, 
            Cermakova K, Hodges HC
  TITLE     beta-catenin functions as a molecular adapter for disordered cBAF 
            interactions.
  JOURNAL   Mol Cell. 2025 Jul 15:S1097-2765(25)00576-3. doi: 
            10.1016/j.molcel.2025.06.026.
  PUBMED    40695292
REFERENCE   2  (bases 1 to 8276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2025 Jul 15:S1097-2765(25)00576-3. doi: 
            10.1016/j.molcel.2025.06.026."
COMMENT     Sequence Label: pPB-CAG-NR5A1-FLAG
FEATURES             Location/Qualifiers
     source          1..8276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     343..362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(581..685)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(711..1166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(848..869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1060..1079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1293..1315
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1307..1324
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1308..1324
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1334..1353
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1334..1352
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   1382..1416
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1737..2116
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3377..3399
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      3490..3499
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4888..4953
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    5048..5597
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5192..5209)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5419..5438
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(6068..6085)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6074..6298
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6345..6361)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6369..6385
                     /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(6393..6399,6400..6417,6418..6423))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6438..6459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   6693..6755
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6796..6816)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6796..6814)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6835..6851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6859..6875
                     /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(6883..6889,6890..6907,6908..6913))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6928..6949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7066..7083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7237..7825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7317..7336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7996..8276,1..511,512..580))
                     /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"
ORIGIN
        1 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
       61 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
      121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
      181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
      241 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
      301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
      361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
      421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
      481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
      541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
      601 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
      661 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
      721 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
      781 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
      841 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
      901 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
      961 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     1021 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     1081 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     1141 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     1201 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     1261 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     1321 cagtgagcgc gcgtaatacg actcactata gggcgaattg gggcgcgcca ttctagatta
     1381 accctagaaa gatagtctgc gtaaaattga cgcatgcatt cttgaaatat tgctctctct
     1441 ttctaaatag cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc gcttggagct
     1501 cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat tttgaactat aacgaccgcg
     1561 tgagtcaaaa tgacgcatga ttatctttta cgtgactttt aagatttaac tcatacgata
     1621 attatattgt tatttcatgt tctacttacg tgataactta ttatatatat attttcttgt
     1681 tatagatatc aactagaatg ctagactcga gaagcttgat cggccacgcg gatctcgaca
     1741 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1801 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1861 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1921 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1981 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2041 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2101 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     2161 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     2221 ggcggggggg gggggggggg gcgccagggg gggcgggggg gggggggggg gggggggggg
     2281 gggggggggg ggggggcggg ggggggcggg ggggggggcg gggcgaggcg gagaggtgcg
     2341 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
     2401 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
     2461 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
     2521 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
     2581 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
     2641 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
     2701 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     2761 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     2821 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     2881 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     2941 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     3001 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc
     3061 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     3121 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     3181 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     3241 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     3301 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     3361 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag ccggagcgca
     3421 cgtcggcagt cggctccctc gttgaccgaa tcaccgacct ctctccccag ggggatccac
     3481 cgggctagcg ccaccatgga ctattcgtac gacgaggacc tggacgagct gtgccccgtg
     3541 tgcggggaca aggtgtccgg ctaccactac ggactgctca cgtgtgagag ctgcaagggc
     3601 ttcttcaagc gcacggtgca gaacaacaag cactacacgt gcaccgagag ccagagctgc
     3661 aagatcgaca agacgcagcg caagcgctgt cccttctgcc gcttccagaa atgcctgacg
     3721 gtggggatgc gcctggaagc cgtgcgcgct gaccgtatga ggggtggccg gaacaagttt
     3781 gggccgatgt acaagcggga ccgggccctg aaacagcaga agaaggcaca gattcgggcc
     3841 aatggcttca agctggagac agggcccccg atgggggtgc ccccgccgcc ccctcccgca
     3901 ccggactacg tgctgcctcc cagcctgcat gggcctgagc ccaagggcct ggccgccggt
     3961 ccacctgctg ggccactggg cgactttggg gccccagcac tgcccatggc cgtgcccggt
     4021 gcccacgggc cactggctgg ctacctctac cctgcctttc ctggccgtgc catcaagtct
     4081 gagtacccgg agccttatgc cagcccccca cagcctgggc tgccgtacgg ctacccagag
     4141 cccttctctg gagggcccaa cgtgcctgag ctcatcctgc agctgctgca gctggagccg
     4201 gatgaggacc aggtgcgggc ccgcatcttg ggctgcctgc aggagcccac caaaagccgc
     4261 cccgaccagc cggcggcctt cggcctcctg tgcagaatgg ccgaccagac cttcatctcc
     4321 atcgtggact gggcacgcag gtgcatggtc ttcaaggagc tggaggtggc cgaccagatg
     4381 acgctgctgc agaactgctg gagcgagctg ctggtgttcg accacatcta ccgccaggtc
     4441 cagcacggca aggagggcag catcctgctg gtcaccgggc aggaggtgga gctgaccaca
     4501 gtggccaccc aggcgggctc gctgctgcac agcctggtgt tgcgggcgca ggagctggtg
     4561 ctgcagctgc ttgcgctgca gctggaccgg caggagtttg tctgcctcaa gttcatcatc
     4621 ctcttcagcc tggatttgaa gttcctgaat aaccacatcc tggtgaaaga cgctcaggag
     4681 aaggccaacg ccgccctgct tgactacacc ctgtgccact acccgcactg cggggacaaa
     4741 ttccagcagc tgctgctgtg cctggtggag gtgcgggccc tgagcatgca ggccaaggag
     4801 tacctgtacc acaagcacct gggcaacgag atgccccgca acaacctgct catcgaaatg
     4861 ctgcaagcca agcagactac cggtatggac tacaaagacc atgacggtga ttataaagat
     4921 catgatatcg attacaagga tgacgatgac aagtgaacgc gttagtgcgg atcctagtgc
     4981 gaattctagt gcgtcgactc tagcaacggt ttccctctag cgggatcaat tccgcccccc
     5041 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     5101 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     5161 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     5221 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     5281 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     5341 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     5401 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     5461 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     5521 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     5581 tgaaaaacac gataatacca tggtcatgaa aacatttaac atttctcaac aagatctaga
     5641 attagtagaa gtagcgacag agaagattac aatgctttat gaggataata aacatcatgt
     5701 gggagcggca attcgtacga aaacaggaga aatcatttcg gcagtacata ttgaagcgta
     5761 tataggacga gtaactgttt gtgcagaagc cattgcgatt ggtagtgcag tttcgaatgg
     5821 acaaaaggat tttgacacga ttgtagctgt tagacaccct tattctgacg aagtagatag
     5881 aagtattcga gtggtaagtc cttgtggtat gtgtagggag ttgatttcag actatgcacc
     5941 agattgtttt gtgttaatag aaatgaatgg caagttagtc aaaactacga ttgaagaact
     6001 cattccactc aaatataccc gaaattaaaa gtcgagatgc atcgatgatc tagagctcgc
     6061 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6121 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6181 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     6241 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     6301 tctgaggcgg aaagaacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     6361 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6421 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6481 gctttccagt cgggaaacct gtcgtgccag cggatcttaa aagttttgtt actttataga
     6541 agaaattttg agtttttgtt tttttttaat aaataaataa acataaataa attgtttgtt
     6601 gaatttatta ttagtatgta agtgtaaata taataaaact taatatctat tcaaattaat
     6661 aaataaacct cgatatacag accgataaaa cacatgcgtc aattttacgc atgattatct
     6721 ttaacgtacg tcacaatatg attatctttc tagggttaat ctagtatcgt agattaatta
     6781 atccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     6841 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6901 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6961 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8101 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgc
//
