LOCUS       sequence-420990-        6603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Msl1+exon(48nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6603)
  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 6603)
  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 Msl1+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    578..699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(615..634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     669..688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(748..764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(748..764)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(761..783)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    772..788
                     /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(796..802,803..820,821..826))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    841..862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(979..996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1150..1738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1230..1249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1909..2700,2701..2769))
                     /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     2532..2551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2770..2874)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2949..2968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3167..3185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3204..3222
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3241..3259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3278..3296
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3318..3342
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3494..3517
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3518..3542
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5394..5418)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5460..5477)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5466..5690
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5823..5842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5974..6021
                     /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)."
     CDS             join(6029..6603,1..448)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
       61 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
      121 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
      181 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
      241 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
      301 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
      361 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
      421 cagcactcgt ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc
      481 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
      541 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
      601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
      661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
      721 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      781 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      841 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      901 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      961 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1021 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1561 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1681 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1741 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1801 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1861 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2041 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2221 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2761 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2821 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2881 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     2941 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     3001 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     3061 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     3121 cgggccagat atacgcgtct cgagggatcc aagctggccg cacgtctccc tatcagtgat
     3181 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtacgt ctagaacgtc
     3241 tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt
     3301 accccctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
     3361 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccgcgg aggctggatc
     3421 ctctagcgtt taaacttaag cttccaccat ggactacaaa gaccatgacg gtgatacata
     3481 tgatcatgac atcgattaca aggatgacga tgacaagaca agtttgtaca aaaaagttgg
     3541 caccaccatg agatccgcag tgttcaaggc ggccgcggcc cctgccggcg gcaaccccga
     3601 gcagcgactg gactacgagc gggctgcggc gctgggcggg cccgaggacg agtccggggc
     3661 ggccgaagcc catttcctcc cccggcatcg taagctcaag gagccggggc ccccgctggc
     3721 ctcttcccag ggcgggagcc cctcgccctc tccagccggc tgcggcggcg gcaagggccg
     3781 gggcttgtta ctcccagccg gggcggcccc cgggcagcag gaagagagct ggggcggttc
     3841 ggtgcccttg ccctgtccgc ccccagctac taaacaagcc ggcatcggcg gggagccagt
     3901 cgcagccggc gctggctgca gcccccggcc caagtatcag gcggtgctgc ccattcagac
     3961 gggctctatc gtggtggcgg cggccaaaga gcctacgccc tgggctgggg acaagggtgg
     4021 ggcggctccc ccagctgcca ccgcctcgga cccggcggga cccccaccac tacctctgcc
     4081 cgggccacca cccctcgcgc ccaccgccac tgctgggacc ctggcggcca gtgagggcag
     4141 atggaagagt ataaggaaga gccctctcgg gggtggcggc ggctcgggag cctccagtca
     4201 ggccgcctgc ctcaaacaga tccttctgct gcaattggac ctcatcgaac agcagcagca
     4261 gcagttgcag gccaaggaga aggagataga ggagctgaag tccgagagag atacgctcct
     4321 tgctcggatt gaacgtatgg aaaggcggat gcagctggtg aagagggata acgagaagga
     4381 aaggcacaag ctgctccagg gctatgaacc tgaagagaga gaggaagcag agttgtctga
     4441 gaaaattaaa ttggagcgcc agccggagct ttgcgagacc tcccaggctc tgccttccaa
     4501 gcctttctca tgtggccgga gtggaaaggg acacaaaagg aaaaccccat ttggaaatac
     4561 agaaagaaag actcctgtta aaaagctggc tcctgaattt tcaaaagtca aaacaaaaac
     4621 tcctaagcac tctcccatta aagaggaacc ctgtggttcc atatcagaaa ctgtttgtaa
     4681 acgtgaattg aggagccaag aaaccccaga aaagccccgg tcttcagtgg ataccccacc
     4741 aagactctcg actccccaaa agggacccag cacccacccc aaggagaaag ccttctcaag
     4801 tgagatggaa gatttgccgt acctttccac cacagaaatg tatttgtgtc gttggcacca
     4861 gcctccccca tcaccgttac cattacggga atcctctcca aagaaggagg agactgtagc
     4921 aaggtgtctg atgccatcaa gtgttgcagg agaaacttca gtcttggctg ttccttcttg
     4981 gagggaccac tcagtagagc ctctaaggga cccaaatcct tcagacattt tggagaacct
     5041 ggatgacagt gtattttcaa agaggcatgc aaaactggag ctagacgaga agaggagaaa
     5101 acgatgggat attcagagga tcagggaaca aagaatttta cagcgactgc agctcagaat
     5161 gtataaaaag aaaggaattc aggaatctga gcctgaggtt acctcatttt tccctgagcc
     5221 agatgatgtt gaaagtttgc tgattacccc cttcttgcct gttgtagcat ttggacggcc
     5281 attgccaaaa ttagctccac agaactttga actaccctgg ttggatgagc gaagccgttg
     5341 caggttggag atccagaaga agcacacacc tcaccggacc tgtaggaagt agtacccaac
     5401 tttcttgtac aaagtggtct gactcgagtc tagagggccc gtttaaaccc gctgatcagc
     5461 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     5521 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     5581 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     5641 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     5701 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     5761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5941 aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc
     6001 tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga
     6061 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     6121 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     6181 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
     6241 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     6301 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     6361 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
     6421 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     6481 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     6541 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     6601 gca
//
