LOCUS       sequence-421022-        6231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Mef2a-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6231)
  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 6231)
  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..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(629..676)
                     /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)."
     primer_bind     808..827
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    960..1184
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1173..1190
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    1232..1256
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2736..2760)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2761..2784)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2936..2960)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2982..3000
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3019..3037
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3056..3074
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3093..3111
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3310..3329
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3404..3508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3509..3577,3578..4369)
                     /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(3727..3746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4540..5128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5029..5048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5282..5299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5416..5437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5452..5457,5458..5475,5476..5482)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5490..5506
                     /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     5495..5517
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5514..5530
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5514..5530
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5579..5700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5590..5609)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5644..5663
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5830..6231,1..621))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg
       61 gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt
      121 gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc
      181 acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg
      241 gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg
      301 cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata
      361 ggtcaggctc tcgctgaact ccccaatgtc aagcacttcc ggaatcggga gcgcggccga
      421 tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg
      481 caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga
      541 gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt
      601 cgcggtgagt tcaggctttt tggccaagga agttcctata ctttctagag aataggaact
      661 tcggaatagg aacttctagg tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
      721 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
      781 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
      841 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
      901 gctacagggc gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc
      961 catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct
     1021 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
     1081 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
     1141 gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg
     1201 ggtttaaacg ggccctctag actcgagtca gaccactttg tacaagaaag ttgggtatta
     1261 ggtcacccat gtgtccatcc tcatgcgttt tacagaaggg ctttctctgt cctcagtatt
     1321 tggaggtcgg ccaagcacaa ttggagaatg gaagtcaccc cgtggatcct cccggtcact
     1381 gccatcatag gagctgctgg agctgctcag actgtccaca ggggagcgcc ccatttcctg
     1441 ccggggttgg ggctgtggag gttgtggcgg tggctgctgt tgtggctgct gctgctgctg
     1501 ttggaagcct gatggggtca ttctatcacg gggaggtgaa attggctctg acttaatgtt
     1561 gatgttctgg ttggtgttaa tggacaaatt tgaaccctga gataactgcc ctccagccac
     1621 cagagagctg agggctgctt gtcctagatg gtgctgctgc caggctgaag cctgccccag
     1681 agacagcatt ccaggggaag taaagccttg cagggcagac aggtcagcgc tggtcagtga
     1741 gtaatcagtg ttgtaggctg tcggcattgc tgagtacact agtccctgtg gaggcaagct
     1801 cggagttgtc acagacacta caggcgtggc aagaggttga gtggcttgag aactgcttat
     1861 cctttgggca ttaagtgggg gcatcatgcc cttgcttgat gggggaatga caactcgtag
     1921 atctggtttc cgactgttca ttccaagact gccaccacct ggtggcgggg gagactttgt
     1981 aggcatgact ttgcctaaac tatttgcacc agtatttcca atcaaatttg gagaggccct
     2041 tgagtttaca aatccattcc ccacaggact atttccagct ccatttggca ccgtgaggtc
     2101 tgtagtgctc aacatcccac ttgcactgcc ggtacttggt ggtctctgcg gagctcctgg
     2161 ggacacattt ctatgtagcg tggctggggg tggagacagc atgctcgaat ctgctaatgt
     2221 tgagctggct gccaaagacg gtgacacgag tgaactccct gggttagtgt acgacaaagc
     2281 attagggctg gtcacaggga ctgtgacaga cattgagaag ttctgaggtg gcaagccagg
     2341 tgcgatttta tgattccgca tcatattatc aaattcctca ttaatttttt tatatttttc
     2401 ttctgtatgt ggagttagca catatgaagt atcagggtct gggctgtcgc accctctgtg
     2461 ttccttcttg ttcagagcct caacgatatc cgagttcgtc ctgctttcat gaggctcgtt
     2521 atactcagtg tatttgagaa ggactttgtc catgtcagtg ctagcgtact gaaacagctt
     2581 attagagctg ttgaaaatga tgagtgctat ttcacagtca cagagcacac tgagttcata
     2641 ggctttcttc atcaatccaa actttcgctt tgtaaaagta acctgtcggt tcctctcatc
     2701 cattatgcgt gtgatttgta tcttctttcg cccggtgcca acttttttgt acaaacttgt
     2761 cttgtcatcg tcatccttgt aatcgatgtc atgatcatat gtatcaccgt catggtcttt
     2821 gtagtccatg gtggaagctt aagtttaaac gctagaggat ccagcctccg cggaggctgg
     2881 atcggtcccg gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc caggcgatct
     2941 gacggttcac taaacgagct ctgcttatat agggggtacc ttctctatca ctgataggga
     3001 gctcgagacg tgtcgacttc tctatcactg atagggagac gttctagacg taccttctct
     3061 atcactgata gggagctcga gacgtgtcga cttctctatc actgataggg agacgtgcgg
     3121 ccagcttgga tccctcgaga cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac
     3181 gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt
     3241 aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc agatactggc
     3301 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga tcgggagatc
     3361 tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3421 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3481 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3541 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3601 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3661 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3721 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3781 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3841 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3901 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3961 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4021 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4081 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4141 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4201 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4261 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4321 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4381 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4441 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4501 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4561 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4621 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4681 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4741 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4801 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4861 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4921 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4981 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5041 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5101 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5161 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5221 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5281 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5341 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5401 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5461 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5521 cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata
     5581 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     5641 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     5701 ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     5761 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     5821 tagtacgtgc tattcctttg ccctcggacg agtgctgggg cgtcggtttc cactatcggc
     5881 gagtacttct acacagccat cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg
     5941 cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc
     6001 atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa tgcggagcat
     6061 atacgcccgg agccgcggcg atcctgcaag ctccggatgc ctccgctcga agtagcgcgt
     6121 ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga cctcgtattg
     6181 ggaatccccg aacatcgcct cgctccagtc aatgaccgct gttatgcggc c
//
