LOCUS       sequence-421000-        8200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Smarcc2-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8200)
  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 8200)
  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..8200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    63..287
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     276..293
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    335..359
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3660..3684)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3685..3708)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3860..3884)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3906..3924
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3943..3961
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3980..3998
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4017..4035
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4054..4072
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4091..4109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4128..4146
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4165..4183
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4382..4401
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4476..4580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4581..4649,4650..5441)
                     /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(4799..4818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5612..6200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6101..6120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6354..6371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6488..6509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6524..6529,6530..6547,6548..6554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6562..6578
                     /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     6567..6589
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6586..6602
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6586..6602
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6651..6772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6662..6681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6716..6735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6902..7924)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(7932..7979)
                     /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     8111..8130
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt ccgcctcaga
       61 agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt
      121 gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc
      181 aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg
      241 cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca
      301 gcgggtttaa acgggccctc tagactcgag tcagaccact ttgtacaaga aagttgggta
      361 tcactgtgga ggtggcacag gggtgactgt gcctgggctt ggagctgtgg ggtctggagg
      421 cagcggggtg cctgggtccg ccacgcctgg gtgcccgctg cctggcactg cccctggcat
      481 cattgaggga ggagttggtt ggttggggaa cggtgagggg ccatggggtc cagggggggg
      541 tacccctggt gggactgccc caggctgagg ggctccagct tgttgtggct gctgtggccc
      601 tgcagttgtc cctgcctgcc caatctgttc agaagggccc aagcttccag gaggggcccc
      661 actgccagga ggggcagagg ccacagcggc tggaggcaca gctagaccat ggactgtagg
      721 tggtccagca gccccggtgg gaggtatggg ctgggagcct gggggcaagg ttggtggctg
      781 ctgctgctgc tgctggtgca tctgctggaa gtgctgctgc cgggccctca tctctgcata
      841 cttcagctgc tccatgtgga aggcttgccg gtcggccagg agctgctgcc tctggtattc
      901 cagcgcctct cgctcccggt ccattattgt ctccagctcc tcaaaatgtc ggagtttgat
      961 ctctagtttc ttcatttggg tctctaccag cagagccacc aaagacttga tctttctctc
     1021 ctcaactgca gccaagtgct tggccttgac tgcagcagcg gccagggctg cggctgcagc
     1081 tgtggacagg ttgccttcac caatgtcacg ctccaccttg gttttcctct ccccctctgg
     1141 ctctgccact tcctttagca cttcctcctg cccttctgtt ggttccttgt ctttctcagg
     1201 atcaactatc gggtccccgt cactcttctc agactccttc tcactgtcac cttctttccc
     1261 tttctcttca tctttcttgg ggacctcact tatttcctcc tttgcttctt cctccacagc
     1321 gccccctcct tcccgtggtt ccttaggctc cttcttctca tctgctgcct ggccctctgg
     1381 ccgtgcctcc tcagtcccgc tttcctcaat gcgctcaggc tcatcagagg cagtccctgc
     1441 gatgccgcta ctctccagac caaaggctgg gtcggccttg cctgtgactt tggccgcttc
     1501 ttcgacctta cgcacgtggg cttccaccaa agctgtgggc acctcttcct tcatttttga
     1561 gaactcttct agggctgact tcgcagcagc agaggcaact cggggatcga cgacagaggc
     1621 caggaaggca acggtgctca taacagggtt gcctgactga ctgaagggga tgggttggta
     1681 ggccagaggg cctagagaag cctccgagtc ctccaggtat gggtcttcaa tgggaaggcg
     1741 gagaaaatgc aagatgcact cgtcctgcgt gcggcttccc acgtgctcag atactttgtt
     1801 ccagtcgtcc ttgtacattt ccaaagcctc caggagcagc agagtctcct gctccgtcca
     1861 ttcccgagtg gcacttgctg cagctttgct cttggagggg acgttcttct ttgtgtacat
     1921 gtctgtgcgc agcccaaaat tctgcatgtc tgctggtttc tccttgccct tctcagggaa
     1981 gttcagcatt tgctgggaag cagagctctg ctgtggaggc ttcggctgaa gaggaaccag
     2041 ccctgatggt gtgtccgcca agacatggaa gtgagaggtg ggtggaggcc ccattggggt
     2101 tggtcggctc tcagcatcta cctggtagtt aataagaccc cactgttcca ggaaggcatg
     2161 gaccctcatg atagcgcaga catcacccgc caaattccgc cgacaggcag tagatgttag
     2221 atactcctgg ggattcagtc ggtaagtgtc aatcatgaag ttccgatacg ccaggtagat
     2281 ctctggagtc ttagacttgt tcttgccgtt gaagaactca ggaagagccc tccgttcaat
     2341 ggcatggacg ctgttgtagt caaaccaggc ggcgtagctg gggatgatga tgtggtgggt
     2401 ctgctcggtc acattgtcct catgcaggtc cgggttcttc gtctgctcgc ctttgttgcc
     2461 cgtgctgttc tcatcctcgt ccttgccggt ggtctccatg ctttcatcgt cctgttcatc
     2521 caggtcagtc atggtgccgc ctttgactgg ggctgactca gagtcctttt tagtgttgac
     2581 tgttttgggg agtgtcacct cttccacgtt tgggactgga gagggctcat ccatgtcttt
     2641 tgtcaggtct tcttgttcct cttctctgtg gcctcgcttt gacttggtat aaggtgttga
     2701 gggtcctttc ttagcgtttt tcttcttagc ctctggggtg ggtgaaggag agggagagcg
     2761 cttcctcttc ttatagttgc cccccttctt gtctcgtctg tctgaatctg ggctgtttac
     2821 ctcgtctgtc agcgtcttgg ctgagatctt cttcctgcgg gagactgggc ttttgtcgtc
     2881 actgacttcg tagtcttcct cattcatcca ctcgttgaat gtgtcggtgt cgaggatcca
     2941 cttcgcatgg accttcctcg gtttctcagg agtgggagcg tcctccacag atgcttcaat
     3001 ttcactcgct gggatccacg tgtcgtagct gtcaggatag tagccccagt gcagaagaac
     3061 ctgtttatcc ctcttcatga ctggccgtac ccattcctct tcttctaggt tccctgggac
     3121 aggatacaca acatgggagg cattgctctt atcctcagag atggttccct ggtgtctctt
     3181 aacaatgtct tttaatttcc ctagcagttt gggctcaatt tctgggcaga ggaaaatgtt
     3241 aggtcgtgac aggcaattat tctgtaccaa ggacttctca atggtcatga acatttccac
     3301 attgcggtcc atgcgtgatg gattctggaa atcgtaacgc cgccatccct ggtcactctt
     3361 gaatttgtag gcagctgcaa gaatatggca gagggatcct cctgctttga aatctaggaa
     3421 acatttgatc ggcagtttag taagcggtgc gttgctgaca tgtttgccaa aaacctcttc
     3481 ctgaaactgg agcaactgca ccaccaggct ggacagagac ttgttggttg gcggttctgc
     3541 ttgtatgtac ttcttgtagt tcttgccgag ccagagccgc acgttgtcga actgggtcac
     3601 ggtgtccgcg gcctcgtagt acttcacgtt ggggccgccg tccttcttcc gcaccgcggt
     3661 gccaactttt ttgtacaaac ttgtcttgtc atcgtcatcc ttgtaatcga tgtcatgatc
     3721 atatgtatca ccgtcatggt ctttgtagtc catggtggaa gcttaagttt aaacgctaga
     3781 ggatccagcc tccgcggagg ctggatcggt cccggtgtct tctatggagg tcaaaacagc
     3841 gtggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt atataggggg
     3901 taccttctct atcactgata gggagctcga gacgtgtcga cttctctatc actgataggg
     3961 agacgttcta gacgtacctt ctctatcact gatagggagc tcgagacgtg tcgacttctc
     4021 tatcactgat agggagacgt tctagacgta ccttctctat cactgatagg gagctcgaga
     4081 cgtgtcgact tctctatcac tgatagggag acgttctaga cgtaccttct ctatcactga
     4141 tagggagctc gagacgtgtc gacttctcta tcactgatag ggagacgtgc ggccagcttg
     4201 gatccctcga gacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc
     4261 ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc ttaaattttg
     4321 ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg gcttaactat
     4381 gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga tctcccgatc
     4441 cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4501 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4561 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4621 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4681 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4741 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4801 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4861 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4921 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4981 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5041 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5101 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5161 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5221 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5281 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5341 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5401 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5461 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5521 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5761 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5821 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6241 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6301 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6361 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6421 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6481 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6541 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6601 accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat
     6661 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     6721 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg
     6781 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     6841 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtagtacgt
     6901 gctattcctt tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt
     6961 ctacacagcc atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag
     7021 tcccggctcc ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga
     7081 aattgccgtc aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc
     7141 ggagccgcgg cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct
     7201 ccatacaagc caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc
     7261 cgaacatcgc ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat
     7321 tgttggagcc gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa
     7381 gcatcagctc atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt
     7441 gccagtgata cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt
     7501 gaccgattcc ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag
     7561 cgatcgcatc catggcctcc gcgaccggct gcagaacagc gggcagttcg gtttcaggca
     7621 ggtcttgcaa cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctga
     7681 actccccaat gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa
     7741 cataacgatc tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc
     7801 ctcctacatc gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg
     7861 agacgctgtc gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct
     7921 ttttggccaa ggaagttcct atactttcta gagaatagga acttcggaat aggaacttct
     7981 aggtacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     8041 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     8101 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     8161 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
//
