LOCUS       sequence-420977-        8219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Smarcc2+exon(93nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8219)
  AUTHORS   Roth JF, Braunschweig U, Wu M, Li JD, Lin ZY, Larsen B, Weatheritt 
            RJ, Gingras AC, Blencowe BJ
  TITLE     Systematic analysis of alternative exon-dependent interactome 
            remodeling reveals multitasking functions of gene regulatory 
            factors.
  JOURNAL   Mol Cell. 2023 Dec 7;83(23):4222-4238.e10. doi: 
            10.1016/j.molcel.2023.10.034.
  PUBMED    38065061
REFERENCE   2  (bases 1 to 8219)
  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: 
            "10.1016/j.molcel.2023.10"; journalName: "Mol Cell"; date: 
            "2023-12-7- 7"; volume: "83"; issue: "23"; pages: "4222-4238.e10"
COMMENT     Sequence Label: 5'-3xFLAG Smarcc2+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..8219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    46..64
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    120..138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     337..356
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        431..535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(536..604,605..1396)
                     /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     complement(754..773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1567..2155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2056..2075
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2309..2326
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2443..2464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2479..2484,2485..2502,2503..2509)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2517..2533
                     /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)."
     primer_bind     2522..2544
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2541..2557
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2541..2557
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2606..2727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2617..2636)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2671..2690
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2857..3879)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(3887..3934)
                     /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)."
     primer_bind     4066..4085
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4218..4442
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4431..4448
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    4490..4514
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7908..7932)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(7933..7956)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(8108..8132)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    8154..8172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8191..8209
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 acgtaccttc tctatcactg atagggagct cgagacgtgt cgacttctct atcactgata
       61 gggagacgtt ctagacgtac cttctctatc actgataggg agctcgagac gtgtcgactt
      121 ctctatcact gatagggaga cgtgcggcca gcttggatcc ctcgagacgc gtatatctgg
      181 cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc atgcaattgt
      241 cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca gcgacctcca
      301 acacacaagc agggagcaga tactggctta actatgcggc atcagagcag attgtactga
      361 gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg
      421 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      481 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      541 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      601 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      661 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      721 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
      781 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
      841 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
      901 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
      961 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1021 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1081 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1141 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1201 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1261 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1321 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1381 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1441 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1501 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1561 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1621 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1681 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     1741 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1801 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1861 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1921 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1981 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2041 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2101 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2161 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2221 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2281 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2341 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     2401 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     2461 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     2521 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct
     2581 agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact
     2641 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2701 accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc
     2761 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     2821 gaaggcggcg gtggaatcga aatctcgtag tacgtgctat tcctttgccc tcggacgagt
     2881 gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc
     2941 cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc
     3001 gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata
     3061 gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc ctgcaagctc
     3121 cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca
     3181 gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat
     3241 gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac
     3301 gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc
     3361 gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc
     3421 aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg
     3481 gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatgg cctccgcgac
     3541 cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc
     3601 acggcgggag atgcaatagg tcaggctctc gctgaactcc ccaatgtcaa gcacttccgg
     3661 aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc
     3721 ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg
     3781 agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag
     3841 aaacttctcg acagacgtcg cggtgagttc aggctttttg gccaaggaag ttcctatact
     3901 ttctagagaa taggaacttc ggaataggaa cttctaggta cgtgaaccat caccctaatc
     3961 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4021 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4081 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4141 cgccgcgctt aatgcgccgc tacagggcgc gtggggatac cccctagagc cccagctggt
     4201 tctttccgcc tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc
     4261 ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact
     4321 cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc
     4381 cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac
     4441 agtcgaggct gatcagcggg tttaaacggg ccctctagac tcgagtcaga ccactttgta
     4501 caagaaagtt gggtatcact gtggaggtgg cacaggggtg actgtgcctg ggcttggagc
     4561 tgtggggtct ggaggcagcg gggtgcctgg gtccgccacg cctgggtgcc cgctgcctgg
     4621 cactgcccct ggcatcattg agggaggagt tggttggttg gggaacggtg aggggccatg
     4681 gggtccaggg gggggtaccc ctggtgggac tgccccaggc tgaggggctc cagcttgttg
     4741 tggctgctgt ggccctgcag ttgtccctgc ctgcccaatc tgttcagaag ggcccaagct
     4801 tccaggaggg gccccactgc caggaggggc agaggccaca gcggctggag gcacagctag
     4861 accatggact gtaggtggtc cagcagcccc ggtgggaggt atgggctggg agcctggggg
     4921 caaggttggt ggctgctgct gctgctgctg gtgcatctgc tggaagtgct gctgccgggc
     4981 cctcatctct gcatacttca gctgctccat gtggaaggct tgccggtcgg ccaggagctg
     5041 ctgcctctgg tattccagcg cctctcgctc ccggtccatt attgtctcca gctcctcaaa
     5101 atgtcggagt ttgatctcta gtttcttcat ttgggtctct accagcagag ccaccaaaga
     5161 cttgatcttt ctctcctcaa ctgcagccaa gtgcttggcc ttgactgcag cagcggccag
     5221 ggctgcggct gcagctgtgg acaggttgcc ttcaccaatg tcacgctcca ccttggtttt
     5281 cctctccccc tctggctctg ccacttcctt tagcacttcc tcctgccctt ctgttggttc
     5341 cttgtctttc tcaggatcaa ctatcgggtc cccgtcactc ttctcagact ccttctcact
     5401 gtcaccttct ttccctttct cttcatcttt cttggggacc tcacttattt cctcctttgc
     5461 ttcttcctcc acagcgcccc ctccttcccg tggttcctta ggctccttct tctcatctgc
     5521 tgcctggccc tctggccgtg cctcctcagt cccgctttcc tcaatgcgct caggctcatc
     5581 agaggcagtc cctgcgatgc cgctactctc cagaccaaag gctgggtcgg ccttgcctgt
     5641 gactttggcc gcttcttcga ccttacgcac gtgggcttcc accaaagctg tgggcacctc
     5701 ttccttcatt tttgagaact cttctagggc tgacttcgca gcagcagagg caactcgggg
     5761 atcgacgaca gaggccagga aggcaacggt gctcataaca gggttgcctg actgactgaa
     5821 ggggatgggt tggtaggcca gagggcctag agaagcctcc gagtcctcca ggtatgggtc
     5881 ttcaatggga aggcggagaa aatgcaagat gcactcgtcc tgcgtgcggc ttcccacgtg
     5941 ctcagatact ttgttccagt cgtccttgta catttccaaa gcctccagga gcagcagagt
     6001 ctcctgctcc gtccattccc gagtggcact tgctgcagct ttgctcttgg aggggacgtt
     6061 cttctttgtg tacatgtctg tgcgcagccc aaaattctgc atgtctgctg gtttctcctt
     6121 gcccttctca gggaagttca gcatttgctg ggaagcagag ctctgcagct ctggcttgcc
     6181 cttagccgtc tctggcacca ggtcatccag ctctttgccc ttccgcccag ccttggtgtc
     6241 agcatcaacc tggcggccct gtggaggctt cggctgaaga ggaaccagcc ctgatggtgt
     6301 gtccgccaag acatggaagt gagaggtggg tggaggcccc attggggttg gtcggctctc
     6361 agcatctacc tggtagttaa taagacccca ctgttccagg aaggcatgga ccctcatgat
     6421 agcgcagaca tcacccgcca aattccgccg acaggcagta gatgttagat actcctgggg
     6481 attcagtcgg taagtgtcaa tcatgaagtt ccgatacgcc aggtagatct ctggagtctt
     6541 agacttgttc ttgccgttga agaactcagg aagagccctc cgttcaatgg catggacgct
     6601 gttgtagtca aaccaggcgg cgtagctggg gatgatgatg tggtgggtct gctcggtcac
     6661 attgtcctca tgcaggtccg ggttcttcgt ctgctcgcct ttgttgcccg tgctgttctc
     6721 atcctcgtcc ttgccggtgg tctccatgct ttcatcgtcc tgttcatcca ggtcagtcat
     6781 ggtgccgcct ttgactgggg ctgactcaga gtccttttta gtgttgactg ttttggggag
     6841 tgtcacctct tccacgtttg ggactggaga gggctcatcc atgtcttttg tcaggtcttc
     6901 ttgttcctct tctctgtggc ctcgctttga cttggtataa ggtgttgagg gtcctttctt
     6961 agcgtttttc ttcttagcct ctggggtggg tgaaggagag ggagagcgct tcctcttctt
     7021 atagttgccc cccttcttgt ctcgtctgtc tgaatctggg ctgtttacct cgtctgtcag
     7081 cgtcttggct gagatcttct tcctgcggga gactgggctt ttgtcgtcac tgacttcgta
     7141 gtcttcctca ttcatccact cgttgaatgt gtcggtgtcg aggatccact tcgcatggac
     7201 cttcctcggt ttctcaggag tgggagcgtc ctccacagat gcttcaattt cactcgctgg
     7261 gatccacgtg tcgtagctgt caggatagta gccccagtgc agaagaacct gtttatccct
     7321 cttcatgact ggccgtaccc attcctcttc ttctaggttc cctgggacag gatacacaac
     7381 atgggaggca ttgctcttat cctcagagat ggttccctgg tgtctcttaa caatgtcttt
     7441 taatttccct agcagtttgg gctcaatttc tgggcagagg aaaatgttag gtcgtgacag
     7501 gcaattattc tgtaccaagg acttctcaat ggtcatgaac atttccacat tgcggtccat
     7561 gcgtgatgga ttctggaaat cgtaacgccg ccatccctgg tcactcttga atttgtaggc
     7621 agctgcaaga atatggcaga gggatcctcc tgctttgaaa tctaggaaac atttgatcgg
     7681 cagtttagta agcggtgcgt tgctgacatg tttgccaaaa acctcttcct gaaactggag
     7741 caactgcacc accaggctgg acagagactt gttggttggc ggttctgctt gtatgtactt
     7801 cttgtagttc ttgccgagcc agagccgcac gttgtcgaac tgggtcacgg tgtccgcggc
     7861 ctcgtagtac ttcacgttgg ggccgccgtc cttcttccgc accgcggtgc caactttttt
     7921 gtacaaactt gtcttgtcat cgtcatcctt gtaatcgatg tcatgatcat atgtatcacc
     7981 gtcatggtct ttgtagtcca tggtggaagc ttaagtttaa acgctagagg atccagcctc
     8041 cgcggaggct ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc
     8101 tccaggcgat ctgacggttc actaaacgag ctctgcttat atagggggta ccttctctat
     8161 cactgatagg gagctcgaga cgtgtcgact tctctatcac tgatagggag acgttctag
//
