LOCUS       sequence-421247-        6276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Mef2d-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6276)
  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 6276)
  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 Mef2d-exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    414..535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(451..470)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     505..524
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(584..600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(584..600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(597..619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    608..624
                     /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(632..638,639..656,657..662))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    677..698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(815..832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(986..1574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1066..1085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1745..2536,2537..2605))
                     /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     2368..2387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2606..2710)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2785..2804)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3003..3021
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3040..3058
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3077..3095
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3114..3132
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3154..3178
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3330..3353
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3354..3378
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4903..4927)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4969..4986)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4975..5199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5332..5351)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5483..5530
                     /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(5538..6276,1..284)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
       61 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
      121 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
      181 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
      241 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
      301 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
      361 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
      421 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
      481 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
      541 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
      601 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      661 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      781 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      841 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      901 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      961 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1021 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1081 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1141 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1201 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1261 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1321 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1381 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1441 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1501 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1561 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1621 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1681 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1741 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1801 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1861 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1921 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1981 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2041 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2101 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2161 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2221 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2281 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2341 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2401 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2461 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2521 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2581 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2641 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2701 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     2761 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2821 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     2881 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     2941 gctgcttcgc gatgtacggg ccagatatac gcgtctcgag ggatccaagc tggccgcacg
     3001 tctccctatc agtgatagag aagtcgacac gtctcgagct ccctatcagt gatagagaag
     3061 gtacgtctag aacgtctccc tatcagtgat agagaagtcg acacgtctcg agctccctat
     3121 cagtgataga gaaggtaccc cctatataag cagagctcgt ttagtgaacc gtcagatcgc
     3181 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     3241 ccgcggaggc tggatcctct agcgtttaaa cttaagcttc caccatggac tacaaagacc
     3301 atgacggtga tacatatgat catgacatcg attacaagga tgacgatgac aagacaagtt
     3361 tgtacaaaaa agttggcacc gggaggaaaa agattcagat ccagcgaatc actgatgaac
     3421 ggaaccgcca ggtgaccttc accaagcgga agtttggact gatgaagaag gcctacgagc
     3481 tgagtgtgct gtgcgactgc gagatcgcgc tcatcatctt caaccactcc aacaagctgt
     3541 tccagtatgc cagcaccgac atggacaagg tgctgctcaa gtacaccgag tacaacgagc
     3601 cacacgagag ccgcaccaat gctgacatca tcgagaccct gaggaagaag ggtttcaacg
     3661 gctgtgacag cccagagccg gatggggagg actcactgga gcagagcccc ctgctggagg
     3721 acaagtaccg gcgggccagt gaggagctgg atgggctctt caggcgctat gggtcatctg
     3781 ttccggcccc caactttgcc atgcctgtca cagtgcccgt gtccaatcag agctccatgc
     3841 agttcagcaa tccaagtagc tctctggtca ctccttccct ggtgacatca tcccttacgg
     3901 acccacggct cctgtccccc cagcagccag cactacagag aaacagtgtt tctccaggct
     3961 tgccccagcg gcctgctagt gcaggagcca tgctgggtgg agacctcaac agtgctaatg
     4021 gagcctgccc cagccccgtt gggaatggct atgtcagtgc ccgagcttcc cctggcctcc
     4081 tccctgtggc caatggcaac agcctaaaca aagtcatccc tgccaagtct ccgcccccac
     4141 ccacccacaa cacccagctt ggagccccca gccgcaagcc tgatctgcgg gtcatcactt
     4201 cccagggagg caaagggtta atgcatcatt tgaacaatgc ccagcgcctt ggggtctccc
     4261 agtctaccca ctcgctcacc accccagtgg tttccgtggc aacaccaagt ttactcagcc
     4321 agggcctccc cttctcctcc atgcccactg cctacaacac agattaccag ctgcccagtg
     4381 cagagctatc ctccttacca gccttcagtt cacctgcagg gctggcacta ggcaatgtca
     4441 ccgcctggca gcagccccag ccgccccagc agccacaacc gccacaaccg ccacagtcac
     4501 agccacagcc accacagcca cagccacagc agccacctca gcaacagccc cacttggtcc
     4561 ccgtttctct cagcaacctc atccctggca gccccttgcc tcacgtgggt gctgctctca
     4621 cagtcactac ccacccccac atcagcatca agtcagaacc agtgtcccca agtcgtgaac
     4681 gcagccctgc acctcctcca ccagctgtgt tcccagctgc ccgccctgag cctggcgaag
     4741 gtctcagcag cccagctgga ggatcctatg agaccgggga ccgggatgat ggacgggggg
     4801 actttgggcc cacactaggc ctgctgcgcc cagccccaga gcctgaggct gagggctcag
     4861 ctgtgaagag gatgcggctg gatacttgga cattaaagtg atacccaact ttcttgtaca
     4921 aagtggtctg actcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     4981 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     5041 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     5101 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     5161 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     5221 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5281 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5341 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5401 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5461 agggtgatgg ttcacgtacc tagaagttcc tattccgaag ttcctattct ctagaaagta
     5521 taggaacttc cttggccaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
     5581 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     5641 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     5701 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     5761 aagtgcttga cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     5821 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
     5881 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     5941 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     6001 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     6061 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     6121 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     6181 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     6241 ggccgtggtt ggcttgtatg gagcagcaga cgcgct
//
