LOCUS       sequence-420983-        6297 bp    DNA     circular SYN 15-APR-2025
DEFINITION  LR vector for integration of Mef2d+exon(21nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  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 6297)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 Mef2d+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    425..546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(462..481)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     516..535
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(595..611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(595..611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(608..630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    619..635
                     /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(643..649,650..667,668..673))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    688..709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(826..843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(997..1585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1077..1096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1756..2547,2548..2616))
                     /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     2379..2398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2617..2721)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2796..2815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3014..3032
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3051..3069
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3088..3106
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3125..3143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3165..3189
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3341..3364
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3365..3389
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4935..4959)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5001..5018)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5007..5231
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5364..5383)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5515..5562
                     /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(5570..6297,1..295)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
       61 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
      121 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
      181 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
      241 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
      301 actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
      361 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
      421 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      481 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
      541 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
      601 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      661 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      721 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      781 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     2761 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2821 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     2881 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     2941 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtctcga gggatccaag
     3001 ctggccgcac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     3061 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     3121 gagctcccta tcagtgatag agaaggtacc ccctatataa gcagagctcg tttagtgaac
     3181 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     3241 cgatccagcc tccgcggagg ctggatcctc tagcgtttaa acttaagctt ccaccatgga
     3301 ctacaaagac catgacggtg atacatatga tcatgacatc gattacaagg atgacgatga
     3361 caagacaagt ttgtacaaaa aagttggcac cgggaggaaa aagattcaga tccagcgaat
     3421 cactgatgaa cggaaccgcc aggtgacctt caccaagcgg aagtttggac tgatgaagaa
     3481 ggcctacgag ctgagtgtgc tgtgcgactg cgagatcgcg ctcatcatct tcaaccactc
     3541 caacaagctg ttccagtatg ccagcaccga catggacaag gtgctgctca agtacaccga
     3601 gtacaacgag ccacacgaga gccgcaccaa tgctgacatc atcgagaccc tgaggaagaa
     3661 gggtttcaac ggctgtgaca gcccagagcc ggatggggag gactcactgg agcagagccc
     3721 cctgctggag gacaagtacc ggcgggccag tgaggagctg gatgggctct tcaggcgcta
     3781 tgggtcatct gttccggccc ccaactttgc catgcctgtc acagtgcccg tgtccaatca
     3841 gagctccatg cagttcagca atccaagtag ctctctggtc actccttccc tggtgacatc
     3901 atcccttacg gacccacggc tcctgtcccc ccagcagcca gcactacaga gaaacagtgt
     3961 ttctccaggc ttgccccagc ggcctgctag tgcaggagcc atgctgggtg gagacctcaa
     4021 cagtgctaat ggagcctgcc ccagccccgt tgggaatggc tatgtcagtg cccgagcttc
     4081 ccctggcctc ctccctgtgg ccaatggcaa cagcctaaac aaagtcatcc ctgccaagtc
     4141 tccgccccca cccacccaca acacccagct tggagccccc agccgcaagc ctgatctgcg
     4201 ggtcatcact tcccagggag gcaaagggtt aatgcatcat ttgactgagg accatttaga
     4261 tctgaacaat gcccagcgcc ttggggtctc ccagtctacc cactcgctca ccaccccagt
     4321 ggtttccgtg gcaacaccaa gtttactcag ccagggcctc cccttctcct ccatgcccac
     4381 tgcctacaac acagattacc agctgcccag tgcagagcta tcctccttac cagccttcag
     4441 ttcacctgca gggctggcac taggcaatgt caccgcctgg cagcagcccc agccgcccca
     4501 gcagccacaa ccgccacaac cgccacagtc acagccacag ccaccacagc cacagccaca
     4561 gcagccacct cagcaacagc cccacttggt ccccgtttct ctcagcaacc tcatccctgg
     4621 cagccccttg cctcacgtgg gtgctgctct cacagtcact acccaccccc acatcagcat
     4681 caagtcagaa ccagtgtccc caagtcgtga acgcagccct gcacctcctc caccagctgt
     4741 gttcccagct gcccgccctg agcctggcga aggtctcagc agcccagctg gaggatccta
     4801 tgagaccggg gaccgggatg atggacgggg ggactttggg cccacactag gcctgctgcg
     4861 cccagcccca gagcctgagg ctgagggctc agctgtgaag aggatgcggc tggatacttg
     4921 gacattaaag tgatacccaa ctttcttgta caaagtggtc tgactcgagt ctagagggcc
     4981 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     5041 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     5101 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     5161 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     5221 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     5281 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5341 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5401 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5461 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt
     5521 cctattccga agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga
     5581 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     5641 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     5701 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     5761 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     5821 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     5881 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     5941 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     6001 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     6061 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     6121 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     6181 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     6241 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagca
//
