LOCUS       sequence-421033-        7329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of L3mbtl3-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7329)
  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 7329)
  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 L3mbtl3-exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..7329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(80..127)
                     /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     259..278
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    411..635
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     624..641
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    683..707
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3285..3309)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3310..3333)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3485..3509)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3531..3549
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3568..3586
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3605..3623
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3642..3660
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3859..3878
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3953..4057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4058..4126,4127..4918)
                     /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(4276..4295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5089..5677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5578..5597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5831..5848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5965..5986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6001..6006,6007..6024,6025..6031)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6039..6055
                     /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     6044..6066
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6063..6079
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6063..6079
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6128..6249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6139..6158)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6193..6212
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6379..7329,1..72))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 caggtcggag acgctgtcga acttttcgat cagaaacttc tcgacagacg tcgcggtgag
       61 ttcaggcttt ttggccaagg aagttcctat actttctaga gaataggaac ttcggaatag
      121 gaacttctag gtacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
      181 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
      241 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
      301 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      361 cgcgtgggga taccccctag agccccagct ggttctttcc gcctcagaag ccatagagcc
      421 caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
      481 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
      541 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
      601 gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc gggtttaaac
      661 gggccctcta gactcgagtc agaccacttt gtacaagaaa gttgggtatc acagttcatt
      721 gtgggaattc ttctccgcag ctttgaacat cagaatagag ttgaaaattt tgagagctgg
      781 gcccagctta atgctcatga ttttaacaat gtctgtttgg gtcatgagta aaaatgcttc
      841 tccatcaatt tgttcatcct taaacacctt tccatgttct tcacacccag gcaagctctg
      901 aatgaattct gacacctcgt ctgtgctcca tttggaaact ctactagctg ggataccagc
      961 gacagtcgga agaagcttgc tctgctgctc ccagcgcaag ggcagacacg gaatgggagg
     1021 cttaaaggag agaaatactg ctgagcgtcg ctgggcctgc tgctgggtct gggcttgctg
     1081 ctgggtgggc tgagcctgtt gaacctgctg aggctgttgg acgggtgggg cctgctgagg
     1141 ctgctgggcc tgctggggct gctgggcctg ctgagcctgt ggagcctgct gggcctgctg
     1201 gacctgtggc gcctgctggg gctgtggagt ctgtggggcc tgtggagcct gctgggcctg
     1261 ctgggcttgc tgggcctgct gggcctgcgg agccttctgt gcctgctgag gaggttgagc
     1321 atgctgcact tgctgcacct gctgaggtgg ctgactctgc tgtacactag gggtaggctc
     1381 ttcccgaaca actttggctt cggctgatgt ctttacttca ttttcttttt tcttttcaga
     1441 atcttctttg aagttattgg catgcttttc cctggtttca ggagacgagg aactttctct
     1501 ggtgtccatc cttttactcc ttgggaacga gggagtagta ggtggagagg tgtcaccact
     1561 taggcggtct gggaagatgc gttccttact caggttgatt tctgaatagg gacagttggc
     1621 accactctga gtgcccaggt atctcgattt tttaaaatga ccaacacccc tgcagcctag
     1681 agttggacat cctccggttt ctgatggttc cattaattct gcagggctta gcggagcctg
     1741 aagagggtgc ccggtcttgg aacaccagcc cacggggtga atatctggag agtctgcatc
     1801 aatccagtag tcatagcaac tgctccagcc atcaaagtga acctttatcc ggtggtcatc
     1861 cgtgtcggct acagttgcta ctctgatgaa cagcgggttt cttttgtcca cagcttccag
     1921 cttcattttc ttctggaatc catgtggagg tttcactttg aaagctcttg caggagcagg
     1981 taaagaattg gtttcttcta aatatttatc ccaggaaaag tgtttcacat gagaatagcc
     2041 tggtggagta atcagggttc ttctgtgctc tttacaccag ccgactggat gaatgtgtgg
     2101 actgtttgat tcacaccaat agtcatagct ctcatcccag ttgtcaaaat gtaccaggaa
     2161 acgattgtcc accatatctg ttaccgtagc aacacagatg acagaagggc tctttttgtc
     2221 tgcagcctcg agcttcatcc caactcgaaa gcctgacggg atcactgtta tgttctgatt
     2281 ttcaaataat gacttgggag cagcctgggc tttgcaagtc ttaaggtagg actgccaatt
     2341 aaattcttct tctttatacc cttttggagg gcggagctta tggccagtct tctcacacca
     2401 gcctactggg tgaatgtcca gagcatctgc atttacccag aagtcatagc aatcagaata
     2461 accgtcaaag tgaagtttta tcctatatcc acaaacctca gccacagtaa ggacgcagta
     2521 cattgcctgg tggtcagggt ccacgccttc tagcttcatg cccactttga atccattttt
     2581 gttatatggg aaggattggt gctctttgaa cagcttcgta ggcacagcca ctgctttctc
     2641 ctcctccagg taggaagccc aacaccaggt cttcttgcct ttaggaggca agccttgctt
     2701 taagacggca gaatccccgc gtcttttcct tctggctctc tgagaacccc tcaggatcct
     2761 tccatcttgt ttgttctcca tttcatcatc tctctcttct ccatcgtcct tggagtccgc
     2821 cttcagagac aattttggct ttttctttcg actacactta ggatcttcct catcgttgtc
     2881 ttcgcctcca tcccgttcat ccttttgatc tttgtctttg gcatgccgag cacaattctg
     2941 gctgcaatac tttcctcccg aaagacattc atctccgttg ccatactgac aacagttctc
     3001 acaaaactga agcccgtctg cttttgattg ctccttcagc ctgaagggta ccccagtctt
     3061 ctctgaaaag acgtcttgag cagttgggac catccatgtg gtagtggctg tagctttctt
     3121 aacactttcc atctcgcttt cgtctgtgat aacttccaaa gctccaaact catttactcg
     3181 aaactttaaa tcactccctg gtagggtacc aacgccatcc ttccagtcca taacactaaa
     3241 cacatcaaac tcttgaccac ttgtgctaga tgcagattca gtggtgccaa cttttttgta
     3301 caaacttgtc ttgtcatcgt catccttgta atcgatgtca tgatcatatg tatcaccgtc
     3361 atggtctttg tagtccatgg tggaagctta agtttaaacg ctagaggatc cagcctccgc
     3421 ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc
     3481 aggcgatctg acggttcact aaacgagctc tgcttatata gggggtacct tctctatcac
     3541 tgatagggag ctcgagacgt gtcgacttct ctatcactga tagggagacg ttctagacgt
     3601 accttctcta tcactgatag ggagctcgag acgtgtcgac ttctctatca ctgataggga
     3661 gacgtgcggc cagcttggat ccctcgagac gcgtatatct ggcccgtaca tcgcgaagca
     3721 gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag ccttgccttg
     3781 ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa gcagggagca
     3841 gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cataggggat
     3901 cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3961 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4021 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4081 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4141 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4201 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4261 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4321 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4381 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4441 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4501 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4561 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4621 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4681 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4741 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4801 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4861 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5221 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5701 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5761 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5821 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5881 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5941 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6001 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6061 cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg acggtataca
     6121 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     6181 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     6241 aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     6301 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     6361 gaaatctcgt agtacgtgct attcctttgc cctcggacga gtgctggggc gtcggtttcc
     6421 actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat
     6481 ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc
     6541 ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat
     6601 gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa
     6661 gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac
     6721 ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc
     6781 attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg
     6841 gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt
     6901 gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc
     6961 acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg
     7021 gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg
     7081 cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata
     7141 ggtcaggctc tcgctgaact ccccaatgtc aagcacttcc ggaatcggga gcgcggccga
     7201 tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg
     7261 caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga
     7321 gagctgcat
//
