LOCUS       sequence-420992-        6555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Msl1-exon into N2a FRT rtTA3 expression
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6555)
  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 6555)
  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..6555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    554..675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(591..610)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     645..664
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(724..740)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(724..740)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(737..759)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    748..764
                     /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(772..778,779..796,797..802))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    817..838
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(955..972)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1126..1714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1206..1225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1885..2676,2677..2745))
                     /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     2508..2527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2746..2850)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2925..2944)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3143..3161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3180..3198
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3217..3235
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3254..3272
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3294..3318
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3470..3493
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3494..3518
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5322..5346)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5388..5405)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5394..5618
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5751..5770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5902..5949
                     /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(5957..6555,1..424)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
       61 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
      121 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
      181 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
      241 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
      301 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
      361 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
      421 atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
      481 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
      541 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
      601 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
      661 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
      721 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      781 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      841 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      961 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1021 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2761 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2821 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2881 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     2941 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     3001 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     3061 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtctcgag
     3121 ggatccaagc tggccgcacg tctccctatc agtgatagag aagtcgacac gtctcgagct
     3181 ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat agagaagtcg
     3241 acacgtctcg agctccctat cagtgataga gaaggtaccc cctatataag cagagctcgt
     3301 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
     3361 caccgggacc gatccagcct ccgcggaggc tggatcctct agcgtttaaa cttaagcttc
     3421 caccatggac tacaaagacc atgacggtga tacatatgat catgacatcg attacaagga
     3481 tgacgatgac aagacaagtt tgtacaaaaa agttggcacc accatgagat ccgcagtgtt
     3541 caaggcggcc gcggcccctg ccggcggcaa ccccgagcag cgactggact acgagcgggc
     3601 tgcggcgctg ggcgggcccg aggacgagtc cggggcggcc gaagcccatt tcctcccccg
     3661 gcatcgtaag ctcaaggagc cggggccccc gctggcctct tcccagggcg ggagcccctc
     3721 gccctctcca gccggctgcg gcggcggcaa gggccggggc ttgttactcc cagccggggc
     3781 ggcccccggg cagcaggaag agagctgggg cggttcggtg cccttgccct gtccgccccc
     3841 agctactaaa caagccggca tcggcgggga gccagtcgca gccggcgctg gctgcagccc
     3901 ccggcccaag tatcaggcgg tgctgcccat tcagacgggc tctatcgtgg tggcggcggc
     3961 caaagagcct acgccctggg ctggggacaa gggtggggcg gctcccccag ctgccaccgc
     4021 ctcggacccg gcgggacccc caccactacc tctgcccggg ccaccacccc tcgcgcccac
     4081 cgccactgct gggaccctgg cggccagtga gggcagatgg aagagtataa ggaagagccc
     4141 tctcgggggt ggcggcggct cgggagcctc cagtcaggcc gcctgcctca aacagatcct
     4201 tctgctgcaa ttggacctca tcgaacagca gcagcagcag ttgcaggcca aggagaagga
     4261 gatagaggag ctgaagtccg agagagatac gctccttgct cggattgaac gtatggaaag
     4321 gcggatgcag ctggtgaaga gggataacga gaaggaaagg cacaagctgc tccagggcta
     4381 tgaacctgaa gagagagagg aagcagagtt gtctgagaaa attaaattgg agcgccagcc
     4441 ggagctttgc gagacctccc aggctctgcc ttccaagcct ttctcatgtg gccggagtgg
     4501 aaagggacac aaaaggaaaa ccccatttgg aaatacagaa agaaagactc ctgttaaaaa
     4561 gctggctcct gaattttcaa aagtcaaaac aaaaactcct aagcactctc ccattaaaga
     4621 ggaaccctgt ggttccatat cagaaactgt ttgtaaacgt gaattgagga gccaagaaac
     4681 cccagaaaag ccccggtctt cagtggatac cccaccaaga ctctcgactc cccaaaaggg
     4741 acccagcacc caccccaagg agaaagcctt ctcaagtgag atggaagatt tgccgtacct
     4801 ttccaccaca gaaatgtatt tgtgtcgttg gcaccagcct cccccatcac cgttaccatt
     4861 acgggaatcc tctccaaaga aggaggagac tgtagcaatt ccttcttgga gggaccactc
     4921 agtagagcct ctaagggacc caaatccttc agacattttg gagaacctgg atgacagtgt
     4981 attttcaaag aggcatgcaa aactggagct agacgagaag aggagaaaac gatgggatat
     5041 tcagaggatc agggaacaaa gaattttaca gcgactgcag ctcagaatgt ataaaaagaa
     5101 aggaattcag gaatctgagc ctgaggttac ctcatttttc cctgagccag atgatgttga
     5161 aagtttgctg attaccccct tcttgcctgt tgtagcattt ggacggccat tgccaaaatt
     5221 agctccacag aactttgaac taccctggtt ggatgagcga agccgttgca ggttggagat
     5281 ccagaagaag cacacacctc accggacctg taggaagtag tacccaactt tcttgtacaa
     5341 agtggtctga ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     5401 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     5461 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     5521 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     5581 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     5641 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5701 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5761 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5821 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5881 gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat
     5941 aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     6001 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     6061 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     6121 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     6181 agtgcttgac attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     6241 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     6301 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     6361 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     6421 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     6481 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     6541 cgcggatttc ggctc
//
