LOCUS       sequence-421020-        6255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Mef2a+exon(24nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6255)
  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 6255)
  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 Mef2a+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    404..525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(441..460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     495..514
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(574..590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(574..590)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(587..609)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    598..614
                     /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(622..628,629..646,647..652))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    667..688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(805..822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(976..1564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1056..1075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1735..2526,2527..2595))
                     /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     2358..2377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2596..2700)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2775..2794)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    2993..3011
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3030..3048
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3067..3085
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3104..3122
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3144..3168
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3320..3343
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3344..3368
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4872..4896)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4938..4955)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4944..5168
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5301..5320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5452..5499
                     /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(5507..6255,1..274)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
       61 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
      121 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
      181 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
      241 acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac
      301 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
      361 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
      421 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
      481 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
      541 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
      601 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      661 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      721 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      781 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      841 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      901 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      961 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1381 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1441 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1501 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1561 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1621 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1681 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1741 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1801 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1861 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1921 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1981 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2041 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2101 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2161 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2221 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2281 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2341 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2401 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2461 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2521 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2581 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2641 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2701 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     2761 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     2821 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     2881 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     2941 gatgtacggg ccagatatac gcgtctcgag ggatccaagc tggccgcacg tctccctatc
     3001 agtgatagag aagtcgacac gtctcgagct ccctatcagt gatagagaag gtacgtctag
     3061 aacgtctccc tatcagtgat agagaagtcg acacgtctcg agctccctat cagtgataga
     3121 gaaggtaccc cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
     3181 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggaggc
     3241 tggatcctct agcgtttaaa cttaagcttc caccatggac tacaaagacc atgacggtga
     3301 tacatatgat catgacatcg attacaagga tgacgatgac aagacaagtt tgtacaaaaa
     3361 agttggcacc gggcgaaaga agatacaaat cacacgcata atggatgaga ggaaccgaca
     3421 ggttactttt acaaagcgaa agtttggatt gatgaagaaa gcctatgaac tcagtgtgct
     3481 ctgtgactgt gaaatagcac tcatcatttt caacagctct aataagctgt ttcagtacgc
     3541 tagcactgac atggacaaag tccttctcaa atacactgag tataacgagc ctcatgaaag
     3601 caggacgaac tcggatatcg ttgaggctct gaacaagaag gaacacagag ggtgcgacag
     3661 cccagaccct gatacttcat atgtgctaac tccacataca gaagaaaaat ataaaaaaat
     3721 taatgaggaa tttgataata tgatgcggaa tcataaaatc gcacctggct tgccacctca
     3781 gaacttctca atgtctgtca cagtccctgt gaccagccct aatgctttgt cgtacactaa
     3841 cccagggagt tcactcgtgt caccgtcttt ggcagccagc tcaacattag cagattcgag
     3901 catgctgtct ccacccccag ccacgctaca tagaaatgtg tccccaggag ctccgcagag
     3961 accaccaagt accggcagtg caagtgggat gttgagcact acagacctca cggtgccaaa
     4021 tggagctgga aatagtcctg tggggaatgg atttgtaaac tcaagggcct ctccaaattt
     4081 gattggaaat actggtgcaa atagtttagg caaagtcatg cctacaaagt ctcccccgcc
     4141 accaggtggt ggcagtcttg gaatgaacag tcggaaacca gatctacgag ttgtcattcc
     4201 cccatcaagc aagggcatga tgcccccact ttcggaggaa gaggaattgg agttgaatgc
     4261 ccaaaggata agcagttctc aagccactca acctcttgcc acgcctgtag tgtctgtgac
     4321 aactccgagc ttgcctccac agggactagt gtactcagca atgccgacag cctacaacac
     4381 tgattactca ctgaccagcg ctgacctgtc tgccctgcaa ggctttactt cccctggaat
     4441 gctgtctctg gggcaggctt cagcctggca gcagcaccat ctaggacaag cagccctcag
     4501 ctctctggtg gctggagggc agttatctca gggttcaaat ttgtccatta acaccaacca
     4561 gaacatcaac attaagtcag agccaatttc acctccccgt gatagaatga ccccatcagg
     4621 cttccaacag cagcagcagc agccacaaca gcagccaccg ccacaacctc cacagcccca
     4681 accccggcag gaaatggggc gctcccctgt ggacagtctg agcagctcca gcagctccta
     4741 tgatggcagt gaccgggagg atccacgggg tgacttccat tctccaattg tgcttggccg
     4801 acctccaaat actgaggaca gagaaagccc ttctgtaaaa cgcatgagga tggacacatg
     4861 ggtgacctaa tacccaactt tcttgtacaa agtggtctga ctcgagtcta gagggcccgt
     4921 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4981 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     5041 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     5101 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     5161 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     5221 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5281 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5341 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5401 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct
     5461 attccgaagt tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact
     5521 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     5581 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     5641 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     5701 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     5761 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     5821 cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga
     5881 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     5941 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     6001 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     6061 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     6121 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     6181 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     6241 gcgctacttc gagcg
//
