LOCUS       sequence-421012-        7773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Med24+exon(57nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7773)
  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 7773)
  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 Med24+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..7773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    189..413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     402..419
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    461..485
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3507..3531)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3532..3555)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3707..3731)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3753..3771
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3790..3808
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3827..3845
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3864..3882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4081..4100
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4175..4279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4280..4348,4349..5140)
                     /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(4498..4517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5311..5899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5800..5819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6053..6070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6187..6208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6223..6228,6229..6246,6247..6253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6261..6277
                     /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     6266..6288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6285..6301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6285..6301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6350..6471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6361..6380)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6415..6434
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6601..7623)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(7631..7678)
                     /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)."
ORIGIN
        1 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
       61 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
      121 taatgcgccg ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc
      181 ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
      241 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
      301 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
      361 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
      421 tgatcagcgg gtttaaacgg gccctctaga ctcgagtcag accactttgt acaagaaagt
      481 tgggtatcag agtgcagcaa tggctttggc ggccacctgc cggcccaggg gcaggctgag
      541 gtctgtgatg gccagaacaa ccttgggact ggacatagca gacactttca ccagttctga
      601 tacggtagtg aagggcatga actgcagaat gctgcccctg ctgtcctgct ccaggcagcc
      661 cacacactcc tccatgaacc actgcacaaa ctgggtgtgg gggccagcgg tgcgtgaacc
      721 caagatggag gaaatgagca ggaacaggtt ggccaggact cggttcagag ggtccctcat
      781 gttgactgtg tggagctggg aggctgagag ggagctgttc atggagcggt cagtgggact
      841 tgacaggatg ttggcatcat cgtcactgga gctcaggagt cgcatcagct tcgagggctg
      901 catgtcctcc acggggaaga ggctgacata gtcctcaatg tcctctctgt gtctcttctt
      961 ctggcgggag gacgcctgtc ccttgtggga agagtaagag ctcagggcac accacacagc
     1021 cagtttggcc agcgcagtgc caggagggtc cataaggctg tgccacttgg aagagtcagt
     1081 aagcagcccg ggcaggatgt gacccagcag gaccagggtc acttgctgca tgtccagaca
     1141 gaagatggag tagagaagct ggaccgccct taacgtgtgc tccttgcgag tctccttcag
     1201 cagctcctta atcaggttgt tgcagaacca gtagacgccg cccatatgca gcagggtgtc
     1261 aaggatgtgg atggagcggc tgtccaccca gcccttctcc agtaccttgg caaagatgtc
     1321 tgtcaggacc tccttgatgg ggcgcttggg aggaagcagg ttccagtagg gcatcgtgtc
     1381 cacgcctgtg gatggaaact tgatctgcgt ggctgtctgc tggagcacgt ccgcacacat
     1441 gtgttccaag attgagttca taatcaccac cctctcattg tagaactgca gggtgttctc
     1501 gctgtacagt ggccctgcca actggcggat catctgcagt gacttctcac gctcatccag
     1561 ccccagcatc cggacgtggg ccacgagcca agccacagca cacaccgcca gactgcatac
     1621 ctttcccttg atattatcgg tgattttctg gatggactca aatgccagaa ccccattctc
     1681 ccaagcgttg aggatttcca aaatggctgc tgaaatgctg aggcaggcct cgtgccactt
     1741 catctgtact agcttcatct ctgaagagtt gttgagcagg gccaccaagg actcgacttt
     1801 ggtggagtca gggcggaagc agggatggtc agggttcaaa atcttgccct cctcgggcat
     1861 gcaggtctgc atccaagtct caaagaaggg gacttcttct cctgagctcg actccgagag
     1921 gatcacctct gagccatagg tctgggccac atggcatagc atgaggaagg aaatgtcaaa
     1981 gagcaaggct cggaccgagg ctgttttcgt gctctctcca ctgccgtggg tcgtgaactc
     2041 attcagattg atgaatttcc gggcgaagga tttaagcttc ccagtggcgg cggcagcagc
     2101 cagcagcaaa tccaggctct tcccagacag catatgtccc aggaccccta gcagtccctc
     2161 cggggacttg gagtgatctg catccattgt cttgagaatg tttgtgaccg tgggctctgc
     2221 ccgcaggatc aggccaggat tcggctggat gttagcattt tctgatgatt ttagctgcgg
     2281 gtcccgatct gccgtgcgtt tgcccacaag gctggcaaag ttgacctcag acagaagccc
     2341 ctgcttgttg cactcctgga ggaggaagtt cgtgcagtcg caattgcagc gctggtcagc
     2401 tttgtccaac aagggggtga gcttcagcag gaactcaaaa gcacagttga catcctccgt
     2461 gaaatccttc tctccatgaa agtacttctt caacttcacc aagacctgtg gaatcttgag
     2521 gtaggtgaaa gctgtccact tcagctcctg cgtgccctcg ggggactcaa tgagtcccac
     2581 aaagcaagct ttccagattt ccaggacaaa gagaggtgtg gggatatgct gcatgcgctt
     2641 caccatcatc agctgctcca ccaggggctg catctccccg gtcaggttca tggtgccctc
     2701 aagcaagatg agtgcgtgga tggtggggaa gccggtcttg tgcagttgct cggagtgcac
     2761 ggacagcatg ctggggatgc tcctgatgag tgtgccgcaa cgctcagctt ggctccggag
     2821 ctgggggttg ctgagattgg ccaggatctc cccaagtttt agaagagaat gctctatggc
     2881 agtccaggag gctgttggtt gattggctcg ggtgccaccc tgcccaagtc cctgggatgt
     2941 atgcaatgag gcctcctcta gtttggcgat gtgtagcagg gctcggttct tggtgctgct
     3001 gagggtcttc tccaggcact gcaagcacaa ggcaagctgc ttctccccgg gggccggagt
     3061 gccagcctcc agcccttcct ggagtcgctc cgcagaagct gctgtacatc gaagcagcca
     3121 gtggagggcg ctaaggagcg ctcggcacaa cccgatgcat tcctctgctt tgccgtggca
     3181 actgagtcgg tcacagaaca tgtccatgat atccagcaga gcctggacac acaggtcccg
     3241 ggaaaagtca tcaaacttac tgatggcggt gagcacagag gagcaggaca ccatctggga
     3301 actaatggcg tacttcaggt acgacaggat cagaggattg ggggaaggcc caatcatagc
     3361 ctgctccagg agcgcttctg ctaggttgag aatgtcccag gtggctcctt tggggaagaa
     3421 ttttttcatg ttgattgccc attggtagtc actccatcgc tctttccagg cttgcaaaat
     3481 ggcctgcttc aggttcacca ccttggtgcc aacttttttg tacaaacttg tcttgtcatc
     3541 gtcatccttg taatcgatgt catgatcata tgtatcaccg tcatggtctt tgtagtccat
     3601 ggtggaagct taagtttaaa cgctagagga tccagcctcc gcggaggctg gatcggtccc
     3661 ggtgtcttct atggaggtca aaacagcgtg gatggcgtct ccaggcgatc tgacggttca
     3721 ctaaacgagc tctgcttata tagggggtac cttctctatc actgataggg agctcgagac
     3781 gtgtcgactt ctctatcact gatagggaga cgttctagac gtaccttctc tatcactgat
     3841 agggagctcg agacgtgtcg acttctctat cactgatagg gagacgtgcg gccagcttgg
     3901 atccctcgag acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc
     3961 taagcagatt cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc
     4021 tcgcgcacta ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg
     4081 cggcatcaga gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc
     4141 gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4201 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4261 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4321 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4381 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4441 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4501 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4561 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4621 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4681 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4741 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4801 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4861 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4921 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4981 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5041 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5101 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5161 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5221 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5281 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5341 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5401 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5461 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5521 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5581 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5641 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5701 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5761 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5821 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5881 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5941 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6001 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6061 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6121 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6181 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6241 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6301 ccatgattac gccaagctct agctagaggt cgacggtata cagacatgat aagatacatt
     6361 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     6421 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt tggggtgggc
     6481 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     6541 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtagtacgtg
     6601 ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc
     6661 tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt
     6721 cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa
     6781 attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg
     6841 gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc
     6901 catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc
     6961 gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt
     7021 gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag
     7081 catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg
     7141 ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg
     7201 accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc
     7261 gatcgcatcc atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag
     7321 gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa
     7381 ctccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac
     7441 ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc
     7501 tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga
     7561 gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt
     7621 tttggccaag gaagttccta tactttctag agaataggaa cttcggaata ggaacttcta
     7681 ggtacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     7741 tcggaaccct aaagggagcc cccgatttag agc
//
