LOCUS       sequence-420972-        8354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Zmynd8-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8354)
  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 8354)
  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 Zmynd8-exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..8354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2508..2532)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2533..2556)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2708..2732)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2754..2772
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2791..2809
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2828..2846
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2865..2883
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2902..2920
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2939..2957
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3156..3175
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3250..3354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3355..3423,3424..4215)
                     /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(3573..3592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4386..4974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4875..4894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5128..5145
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5262..5283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5298..5303,5304..5321,5322..5328)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5336..5352
                     /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     5341..5363
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5360..5376
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5360..5376
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5425..5546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5436..5455)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5490..5509
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5676..6698)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(6706..6753)
                     /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     6885..6904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    7037..7261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7250..7267
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    7309..7333
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 taagagcggc ctctgcttct tcaccgggct tcccgtgacg gcggtggccg gtgccgtgac
       61 tgtgacggta gacatcgtgc tggtggtggc ggcggccacg gtgaccccag ccgcgggtgc
      121 ttgggtggct gagcgggtga gtaccggtgt ctctggggga gactggttgc ccgccgaagt
      181 ggacgacggt ggcggtttac ctatagcgtc ttgcttaggc gacggcacct tgggatcttt
      241 cttgtttttt cgaccctccc tcccagaagg atcctctcct aagtctatga caagttcgct
      301 ctccgagtcc gaatccaagc ccaagtgcac tgtgggagaa tccgtttcat cctttccttt
      361 cagtttgtcc tttgtgggat gaggtgatgg ctttgctttc tctacaaagt ccttctcagg
      421 ctctgggctg gccttgtcct tggcgggtgt ggggtccggc ttctcgctga caggagagtt
      481 gctcttcgct tcctctttga cctctacctg gtttgtgggt ttgggcttcc ttttgatggc
      541 aggggcctct ttgtccaccc gactcccctc tttgtcctcg gggtcctcgg gctcattctt
      601 gggcttctgt tcctcatcgc tgacgtactc gctgtcgctg ctctccgact tttcggagtc
      661 ctcggaatcg ctgtgctcca cggctgtata aacatcctct gagatctcat ttatgcctag
      721 ctgtgctttg caactctcta tggtcttgtc caggttgagc tggaaccggc ttcgaatctg
      781 ccgcttggga gagatgagag ggacgggggc tgactgctgc tggaccgact cgctcagctc
      841 cttcaggtcc atctcggcct tgcttcgatc caagttcagg ttcaggatgc tcccggtggt
      901 ggaggtggaa gtcttgtcgg gttttgtcat gatgggtgtc ggcgcttgcg gagagaaagg
      961 ctttgggctg ccagacaagc tcccggcttg ccccgtcttg gtggaggctg gagaagctgt
     1021 gctcttatca aggaagtcca tggactcctc gcttgcgctg aagtggctgg acggggcctt
     1081 cttctcgggg tcctgctcca catcggagcc cgtgtgcacg gaagagttcg tactggtggg
     1141 ggagcgaggc atgtcggaca gggagatcct gcggccggca cccccgctca gaaggggctt
     1201 gctcagaagg atcttggggg acgctgtcat gtcaaaatta agcttcacct tctcctgttt
     1261 gtctgtcttg gctgtgcccg cgctggggtt gctgggatcc agcagcattt ggtactggtt
     1321 gttgggcgtg tagggcgtcc tgaacgggaa gtaattaaaa accccaaact tcctccgtat
     1381 gttctccacg taaacttcca tctcttgcat ggcgctgttg aagatacttt tagtcttttt
     1441 cacagaaaag gggatttctt tagacatgag gtagcaatta ttgactggaa cccaggctct
     1501 gtcatgttgt ccaaagaaac gggcgtcaac ctgcccgtct ttgtccctca gagctttcgc
     1561 tggccagaat ggaaatcctt tcagttttgc ccagaccaaa gggtgcggat tgctacaggg
     1621 ctcacagaac cagttgtctc gtttttggca agctgcaaga taacattctg gacagacttc
     1681 gatttcattc atctcgtgct cacagatttt gatgacgact tttgctattt gcgtcaactt
     1741 gtgatttccc ccattataaa taatgcagtt gtgcaggatc cacttggcat cggccaggaa
     1801 ggcttctgtg cagccgtaca tcttcttttt tgcattcttt tccaatgtac aaaggtccat
     1861 ggggtggaaa atatattctg catagtcagg gtgttgctcc aatggaacag gcttctggaa
     1921 tgcatccgtc cctggctgct tcattttctg aatggcaaac ttgagcaggt aggacagttg
     1981 ttcaatggtc agcatggtca tggctttgct ctgcgtctcg atgcattctg ctactgtaat
     2041 cttctcacat tcaggacaaa accagtcccc ctctggctcc gatgtcagtc tcagacactt
     2101 agcgtgataa acccggggac agagctcaca gcaaaggact tgtccttccc ggtgacaaac
     2161 ccagcaatag aagtcattcc gtccgtcctg cggtacaaca tcaacagggt ctgtggtgag
     2221 tggctgcttc atatagtaaa acggaccatg tcttagctct gactgttcct tattgctact
     2281 gttgagtaag ccgggtttct ttttcttttt aatggggctc gtggatgagt cttggggcga
     2341 atggccattg gaggaatgtg gagggctggg gaacttccgt ttcggggccg ttttctctgt
     2401 agagacagga tcttccttag gttctttgga gcgagtagag atatccattc cctccaccac
     2461 ctcctgctca gtttttattt cctcttcagc caagctctgt ggatgggtgc caactttttt
     2521 gtacaaactt gtcttgtcat cgtcatcctt gtaatcgatg tcatgatcat atgtatcacc
     2581 gtcatggtct ttgtagtcca tggtggaagc ttaagtttaa acgctagagg atccagcctc
     2641 cgcggaggct ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc
     2701 tccaggcgat ctgacggttc actaaacgag ctctgcttat atagggggta ccttctctat
     2761 cactgatagg gagctcgaga cgtgtcgact tctctatcac tgatagggag acgttctaga
     2821 cgtaccttct ctatcactga tagggagctc gagacgtgtc gacttctcta tcactgatag
     2881 ggagacgttc tagacgtacc ttctctatca ctgataggga gctcgagacg tgtcgacttc
     2941 tctatcactg atagggagac gtgcggccag cttggatccc tcgagacgcg tatatctggc
     3001 ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc agattcttca tgcaattgtc
     3061 ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa
     3121 cacacaagca gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     3181 agtgcaccat aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg
     3241 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3301 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3361 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3421 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3481 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3541 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3601 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3661 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3721 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3781 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3841 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3901 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3961 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4021 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4081 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4141 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4201 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4261 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4321 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4381 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4441 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4501 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4561 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4621 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4681 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4741 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4801 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4861 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4921 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4981 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5041 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5101 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5161 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     5221 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     5281 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     5341 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     5401 gaggtcgacg gtatacagac atgataagat acattgatga gtttggacaa accacaacta
     5461 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     5521 ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg
     5581 cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag
     5641 aaggcggcgg tggaatcgaa atctcgtagt acgtgctatt cctttgccct cggacgagtg
     5701 ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc
     5761 gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg
     5821 tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag
     5881 agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc
     5941 ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag
     6001 aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg
     6061 accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg
     6121 aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg
     6181 acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca
     6241 atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg
     6301 ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc
     6361 ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca
     6421 cggcgggaga tgcaataggt caggctctcg ctgaactccc caatgtcaag cacttccgga
     6481 atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg
     6541 gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga
     6601 gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga
     6661 aacttctcga cagacgtcgc ggtgagttca ggctttttgg ccaaggaagt tcctatactt
     6721 tctagagaat aggaacttcg gaataggaac ttctaggtac gtgaaccatc accctaatca
     6781 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6841 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6901 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6961 gccgcgctta atgcgccgct acagggcgcg tggggatacc ccctagagcc ccagctggtt
     7021 ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc
     7081 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     7141 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     7201 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     7261 gtcgaggctg atcagcgggt ttaaacgggc cctctagact cgagtcagac cactttgtac
     7321 aagaaagttg ggtactagtc ccagaaggca tctagccgag actcccggga ctctttgggc
     7381 atgaggttct tcgtgctggt ccctccactg tgctcagagc gggatgacgc tcgcttctcc
     7441 tcgctgctgc tccacaaacc tgccttgctg ctccgggaat ggtcagagct ttgattggag
     7501 cctaagagca tagaggaggg cgtctctctg gagccggaaa gatccagggt cgagttacta
     7561 ctgtccttgc tcttctccgc tggcgcctct ttctctttgg gggcgctggc cggttctgaa
     7621 ggtgctgact gtgtgttgga ggagttgccc tgcgatgact tatttcctgt ttctgtgctt
     7681 gcctcggcat ctgcttcctg ctgaggggca gtcgccgact gggtacagga cttcatgtgc
     7741 tcgggccagt gggcctgctg acaggggtag tcacagtagc tggtgttcca gcagcagtag
     7801 aaaatggcct ccttcttgca gttggcacac cactgcttct tcttggtctc gtccaccgcc
     7861 tgctgcttct ccagctccag ttgcttcttc acctcggcga tgagccgatc ccgctcctgt
     7921 tccaggctct gccgcatctc ggccatggtt aactccaggt tgtgcttcat ctcagcaagc
     7981 tcctgctggt gcagccactg cagtttctca atctcaatcc tcagccttcg aatctcagct
     8041 attgtgctcc cagtggtgtt cttggagagg tcattgtaga tttctgtcat cgtccccttt
     8101 atggcatcca tcattttgct ggtgtacttg gcaatgtctg cggccacgtc ggccgaggcg
     8161 gtggcaatgg gaaggtcggc attgattgca gacgccaggg tgcttgctga gcccgaactg
     8221 ctgaccaggg ctgccggctg tgtgctggtc accagcgtga tggtggacgt gggtgggctc
     8281 tgggtgacct ccttctgctg gacagttgat gcgttccaca cgacccgctg cacagctggg
     8341 acagtctcct tcgg
//
