LOCUS       sequence-421025-        7716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Med24-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7716)
  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 7716)
  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..7716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1134..1255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1171..1190)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1225..1244
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1304..1320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1304..1320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1317..1339)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1328..1344
                     /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(1352..1358,1359..1376,1377..1382))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1397..1418
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1535..1552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1706..2294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1786..1805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2465..3256,3257..3325))
                     /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     3088..3107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3326..3430)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3505..3524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3723..3741
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3760..3778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3797..3815
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3834..3852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3874..3898
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4050..4073
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    4074..4098
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7063..7087)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7129..7146)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7135..7359
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7492..7511)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    7643..7690
                     /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(7698..7716,1..1004)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
       61 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
      121 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
      181 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga
      241 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
      301 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
      361 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
      421 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
      481 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
      541 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
      601 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
      661 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
      721 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
      781 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
      841 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
      901 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
      961 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
     1021 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     1081 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     1141 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1201 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1261 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     1321 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     1381 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     1441 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1501 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3421 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     3481 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3541 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     3601 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     3661 gctgcttcgc gatgtacggg ccagatatac gcgtctcgag ggatccaagc tggccgcacg
     3721 tctccctatc agtgatagag aagtcgacac gtctcgagct ccctatcagt gatagagaag
     3781 gtacgtctag aacgtctccc tatcagtgat agagaagtcg acacgtctcg agctccctat
     3841 cagtgataga gaaggtaccc cctatataag cagagctcgt ttagtgaacc gtcagatcgc
     3901 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     3961 ccgcggaggc tggatcctct agcgtttaaa cttaagcttc caccatggac tacaaagacc
     4021 atgacggtga tacatatgat catgacatcg attacaagga tgacgatgac aagacaagtt
     4081 tgtacaaaaa agttggcacc aaggtggtga acctgaagca ggccattttg caagcctgga
     4141 aagagcgatg gagtgactac caatgggcaa tcaacatgaa aaaattcttc cccaaaggag
     4201 ccacctggga cattctcaac ctagcagaag cgctcctgga gcaggctatg attgggcctt
     4261 cccccaatcc tctgatcctg tcgtacctga agtacgccat tagttcccag atggtgtcct
     4321 gctcctctgt gctcaccgcc atcagtaagt ttgatgactt ttcccgggac ctgtgtgtcc
     4381 aggctctgct ggatatcatg gacatgttct gtgaccgact cagttgccac ggcaaagcag
     4441 aggaatgcat cgggttgtgc cgagcgctcc ttagcgccct ccactggctg cttcgatgta
     4501 cagcagcttc tgcggagcga ctccaggaag ggctggaggc tggcactccg gcccccgggg
     4561 agaagcagct tgccttgtgc ttgcagtgcc tggagaagac cctcagcagc accaagaacc
     4621 gagccctgct acacatcgcc aaactagagg aggcctcctc ctggactgcc atagagcatt
     4681 ctcttctaaa acttggggag atcctggcca atctcagcaa cccccagctc cggagccaag
     4741 ctgagcgttg cggcacactc atcaggagca tccccagcat gctgtccgtg cactccgagc
     4801 aactgcacaa gaccggcttc cccaccatcc acgcactcat cttgcttgag ggcaccatga
     4861 acctgaccgg ggagatgcag cccctggtgg agcagctgat gatggtgaag cgcatgcagc
     4921 atatccccac acctctcttt gtcctggaaa tctggaaagc ttgctttgtg ggactcattg
     4981 agtcccccga gggcacgcag gagctgaagt ggacagcttt cacctacctc aagattccac
     5041 aggtcttggt gaagttgaag aagtactttc atggagagaa ggatttcacg gaggatgtca
     5101 actgtgcttt tgagttcctg ctgaagctca cccccttgtt ggacaaagct gaccagcgct
     5161 gcaattgcga ctgcacgaac ttcctcctcc aggagtgcaa caagcagggg cttctgtctg
     5221 aggtcaactt tgccagcctt gtgggcaaac gcacggcaga tcgggacccg cagctaaaat
     5281 catcagaaaa tgctaacatc cagccgaatc ctggcctgat cctgcgggca gagcccacgg
     5341 tcacaaacat tctcaagaca atggatgcag atcactccaa gtccccggag ggactgctag
     5401 gggtcctggg acatatgctg tctgggaaga gcctggattt gctgctggct gctgccgccg
     5461 ccactgggaa gcttaaatcc ttcgcccgga aattcatcaa tctgaatgag ttcacgaccc
     5521 acggcagtgg agagagcacg aaaacagcct cggtccgagc cttgctcttt gacatttcct
     5581 tcctcatgct atgccatgtg gcccagacct atggctcaga ggtgatcctc tcggagtcga
     5641 gctcaggaga agaagtcccc ttctttgaga cttggatgca gacctgcatg cccgaggagg
     5701 gcaagatttt gaaccctgac catccctgct tccgccctga ctccaccaaa gtcgagtcct
     5761 tggtggccct gctcaacaac tcttcagaga tgaagctagt acagatgaag tggcacgagg
     5821 cctgcctcag catttcagca gccattttgg aaatcctcaa cgcttgggag aatggggttc
     5881 tggcatttga gtccatccag aaaatcaccg ataatatcaa gggaaaggta tgcagtctgg
     5941 cggtgtgtgc tgtggcttgg ctcgtggccc acgtccggat gctggggctg gatgagcgtg
     6001 agaagtcact gcagatgatc cgccagttgg cagggccact gtacagcgag aacaccctgc
     6061 agttctacaa tgagagggtg gtgattatga actcaatctt ggaacacatg tgtgcggacg
     6121 tgctccagca gacagccacg cagatcaagt ttccatccac aggcgtggac acgatgccct
     6181 actggaacct gcttcctccc aagcgcccca tcaaggaggt cctgacagac atctttgcca
     6241 aggtactgga gaagggctgg gtggacagcc gctccatcca catccttgac accctgctgc
     6301 atatgggcgg cgtctactgg ttctgcaaca acctgattaa ggagctgctg aaggagactc
     6361 gcaaggagca cacgttaagg gcggtccagc ttctctactc catcttctgt ctggacatgc
     6421 agcaagtgac cctggtcctg ctgggtcaca tcctgcccgg gctgcttact gactcttcca
     6481 agtggcacag ccttatggac cctcctggca ctgcgctggc caaactggct gtgtggtgtg
     6541 ccctgagctc ttactcttcc cacaagggac aggcgtcctc ccgccagaag aagagacaca
     6601 gagaggacat tgaggactat gtcagcctct tccccgtgga ggacatgcag ccctcgaagc
     6661 tgatgcgact cctgagctcc agtgacgatg atgccaacat cctgtcaagt cccactgacc
     6721 gctccatgaa cagctccctc tcagcctccc agctccacac agtcaacatg agggaccctc
     6781 tgaaccgagt cctggccaac ctgttcctgc tcatttcctc catcttgggt tcacgcaccg
     6841 ctggccccca cacccagttt gtgcagtggt tcatggagga gtgtgtgggc tgcctggagc
     6901 aggacagcag gggcagcatt ctgcagttca tgcccttcac taccgtatca gaactggtga
     6961 aagtgtctgc tatgtccagt cccaaggttg ttctggccat cacagacctc agcctgcccc
     7021 tgggccggca ggtggccgcc aaagccattg ctgcactctg atacccaact ttcttgtaca
     7081 aagtggtctg actcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     7141 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     7201 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     7261 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     7321 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     7381 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     7441 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     7501 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     7561 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     7621 agggtgatgg ttcacgtacc tagaagttcc tattccgaag ttcctattct ctagaaagta
     7681 taggaacttc cttggccaaa aagcctgaac tcaccg
//
