LOCUS       sequence-421242-        7404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of L3mbtl3+exon(75nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7404)
  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 7404)
  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..7404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(42..89)
                     /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     221..240
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    373..597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     586..603
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    645..669
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3322..3346)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3347..3370)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3522..3546)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     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"
     protein_bind    3679..3697
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3896..3915
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3990..4094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4095..4163,4164..4955)
                     /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(4313..4332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5126..5714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5615..5634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5868..5885
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6002..6023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6038..6043,6044..6061,6062..6068)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6076..6092
                     /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     6081..6103
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6100..6116
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6100..6116
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6165..6286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6176..6195)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6230..6249
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6416..7404,1..34))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tctcgacaga cgtcgcggtg agttcaggct ttttggccaa ggaagttcct atactttcta
       61 gagaatagga acttcggaat aggaacttct aggtacgtga accatcaccc taatcaagtt
      121 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
      181 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
      241 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
      301 cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt
      361 ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
      421 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
      481 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
      541 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
      601 aggctgatca gcgggtttaa acgggccctc tagactcgag tcagaccact ttgtacaaga
      661 aagttgggta tcacagttca ttgtgggaat tcttctccgc agctttgaac atcagaatag
      721 agttgaaaat tttgagagct gggcccagct taatgctcat gattttaaca atgtctgttt
      781 gggtcatgag taaaaatgct tctccatcaa tttgttcatc cttaaacacc tttccatgtt
      841 cttcacaccc aggcaagctc tgaatgaatt ctgacacctc gtctgtgctc catttggaaa
      901 ctctactagc tgggatacca gcgacagtcg gaagaagctt gctctgctgc tcccagcgca
      961 agggcagaca cggaatggga ggcttaaagg agagaaatac tgctgagcgt cgctgggcct
     1021 gctgctgggt ctgggcttgc tgctgggtgg gctgagcctg ttgaacctgc tgaggctgtt
     1081 ggacgggtgg ggcctgctga ggctgctggg cctgctgggg ctgctgggcc tgctgagcct
     1141 gtggagcctg ctgggcctgc tggacctgtg gcgcctgctg gggctgtgga gtctgtgggg
     1201 cctgtggagc ctgctgggcc tgctgggctt gctgggcctg ctgggcctgc ggagccttct
     1261 gtgcctgctg aggaggttga gcatgctgca cttgctgcac ctgctgaggt ggctgactct
     1321 gctgtacact aggggtaggc tcttcccgaa caactttggc ttcggctgat gtctttactt
     1381 cattttcttt tttcttttca gaatcttctt tgaagttatt ggcatgcttt tccctggttt
     1441 caggagacga ggaactttct ctggtgtcca tccttttact ccttgggaac gagggagtag
     1501 taggtggaga ggtgtcacca cttaggcggt ctgggaagat gcgttcctta ctcaggttga
     1561 tttctgaata gggacagttg gcaccactct gagtgcccag gtatctcgat tttttaaaat
     1621 gaccaacacc cctgcagcct agagttggac atcctccggt ttctgatggt tccattaatt
     1681 ctgcagggct tagcggagcc tgaagagggt gcccggtctt ggaacaccag cccacggggt
     1741 gaatatctgg agagtctgca tcaatccagt agtcatagca actgctccag ccatcaaagt
     1801 gaacctttat ccggtggtca tccgtgtcgg ctacagttgc tactctgatg aacagcgggt
     1861 ttcttttgtc cacagcttcc agcttcattt tcttctggaa tccatgtgga ggtttcactt
     1921 tgaaagctct tgcaggagca ggtaaagaat tggtttcttc taaatattta tcccaggaaa
     1981 agtgtttcac atgagaatag cctggtggag taatcagggt tcttctgtgc tctttacacc
     2041 agccgactgg atgaatgtgt ggactgtttg attcacacca atagtcatag ctctcatccc
     2101 agttgtcaaa atgtaccagg aaacgattgt ccaccatatc tgttaccgta gcaacacaga
     2161 tgacagaagg gctctttttg tctgcagcct cgagcttcat cccaactcga aagcctgacg
     2221 ggatcactgt tatgttctga ttttcaaata atgacttggg agcagcctgg gctttgcaag
     2281 tcttaaggta ggactgccaa ttaaattctt cttctttata cccttttgga gggcggagct
     2341 tatggccagt cttctcacac cagcctactg ggtgaatgtc cagagcatct gcatttaccc
     2401 agaagtcata gcaatcagaa taaccgtcaa agtgaagttt tatcctatat ccacaaacct
     2461 cagccacagt aaggacgcag tacattgcct ggtggtcagg gtccacgcct tctagcttca
     2521 tgcccacttt gaatccattt ttgttatatg ggaaggattg gtgctctttg aacagcttcg
     2581 taggcacagc cactgctttc tcctcctcca ggtaggaagc ccaacaccag gtcttcttgc
     2641 ctttaggagg caagccttgc tttaagacgg cagaatcccc gcgtcttttc cttctggctc
     2701 tctgagaacc cctcaggatc cttccatctt gtttgttctc catttcatca tctctctctt
     2761 ctccatcgtc cttggagtcc gccttcagag acaattttgg ctttttcttt cgactacact
     2821 taggatcttc ctcatcgttg tcttcgcctc catcccgttc atccttttga tctttgtctt
     2881 tggcatgccg agcacaattc tggctgcaat actttcctcc cgaaagacat tcatctccgt
     2941 tgccatactg acaacagttc tcacaaaact gaagcccgtc tgcttttgat tgctccttca
     3001 gcctgaaggg taccccagtc ttctctgaaa agactgtggg acatccaggt ggagatgtcc
     3061 agtaggcagg tggaaatacg ggcctggagc tcgggggaga agtcggggcg tcttgagcag
     3121 ttgggaccat ccatgtggta gtggctgtag ctttcttaac actttccatc tcgctttcgt
     3181 ctgtgataac ttccaaagct ccaaactcat ttactcgaaa ctttaaatca ctccctggta
     3241 gggtaccaac gccatccttc cagtccataa cactaaacac atcaaactct tgaccacttg
     3301 tgctagatgc agattcagtg gtgccaactt ttttgtacaa acttgtcttg tcatcgtcat
     3361 ccttgtaatc gatgtcatga tcatatgtat caccgtcatg gtctttgtag tccatggtgg
     3421 aagcttaagt ttaaacgcta gaggatccag cctccgcgga ggctggatcg gtcccggtgt
     3481 cttctatgga ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa
     3541 cgagctctgc ttatataggg ggtaccttct ctatcactga tagggagctc gagacgtgtc
     3601 gacttctcta tcactgatag ggagacgttc tagacgtacc ttctctatca ctgataggga
     3661 gctcgagacg tgtcgacttc tctatcactg atagggagac gtgcggccag cttggatccc
     3721 tcgagacgcg tatatctggc ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc
     3781 agattcttca tgcaattgtc ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg
     3841 cactactcag cgacctccaa cacacaagca gggagcagat actggcttaa ctatgcggca
     3901 tcagagcaga ttgtactgag agtgcaccat aggggatcgg gagatctccc gatccgtcga
     3961 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4021 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4081 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4141 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4201 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4261 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4321 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4381 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4441 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4501 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4561 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4621 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4681 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4921 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5101 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5161 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5221 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5281 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5341 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5401 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5461 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5521 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5581 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5641 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5701 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5761 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5821 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5881 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5941 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6001 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6061 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     6121 attacgccaa gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga
     6181 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     6241 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga
     6301 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     6361 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtagt acgtgctatt
     6421 cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac
     6481 agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg
     6541 ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc
     6601 cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc
     6661 gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac
     6721 aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca
     6781 tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg
     6841 agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca
     6901 gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt
     6961 gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga
     7021 ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg
     7081 catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt
     7141 gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaactccc
     7201 caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac
     7261 gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta
     7321 catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc
     7381 tgtcgaactt ttcgatcaga aact
//
