LOCUS       sequence-421029-        5364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Chtop-exon_siResist into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5364)
  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 5364)
  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 Chtop-exon_siResist minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..5364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(32..1054)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(1062..1109)
                     /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     1241..1260
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1393..1617
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1606..1623
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    1665..1689
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2302..2326)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2327..2350)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2502..2526)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2548..2566
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2585..2603
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2622..2640
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2659..2677
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2876..2895
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2970..3074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3075..3143,3144..3935)
                     /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(3293..3312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4106..4694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4595..4614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4848..4865
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4982..5003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5018..5023,5024..5041,5042..5048)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5056..5072
                     /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     5061..5083
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5080..5096
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5080..5096
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5145..5266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5156..5175)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5210..5229
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cggcggtgga atcgaaatct cgtagtacgt gctattcctt tgccctcgga cgagtgctgg
       61 ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc
      121 ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc
      181 atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt
      241 ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca agctccggat
      301 gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga
      361 agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg
      421 ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt
      481 gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg
      541 acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg
      601 cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga
      661 acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc gcgaccggct
      721 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc
      781 gggagatgca ataggtcagg ctctcgctga actccccaat gtcaagcact tccggaatcg
      841 ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc
      901 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt
      961 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact
     1021 tctcgacaga cgtcgcggtg agttcaggct ttttggccaa ggaagttcct atactttcta
     1081 gagaatagga acttcggaat aggaacttct aggtacgtga accatcaccc taatcaagtt
     1141 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     1201 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     1261 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     1321 cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt
     1381 ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
     1441 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
     1501 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     1561 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     1621 aggctgatca gcgggtttaa acgggccctc tagactcgag tcagaccact ttgtacaaga
     1681 aagttgggta tcaatcattg gtttcaggat ctgtctgtgc catgtaggca tccaattcag
     1741 catccaggtg tcctttggtc ttggacatat aggcatccaa ttggttgtcc agctgctcct
     1801 tggtcaatac agggcgagtg agggcacctc tccctcggcc acgacctcgg cctctgcctc
     1861 caaagccccc tcttccccga cctatcatac cccgacctcg gagcgacatc ccacccctaa
     1921 gcagggttct ggtagcacgt cccccacgta gtcctcctcg gggcaagcct ctctggatta
     1981 tgggtaggcc tcttcctcca attgctcccc tggccagggc acctatgggt cggcctaacc
     2041 gtgcctggat attactctta cccaggcgct gctttaagct cttctgctta agttttaatg
     2101 ctgcctggac agagggtcta ttctccatct gctgggccag tcttctgttt ctggcactgg
     2161 ctagctgctg ctgctgctgc atcgaagccc gaatattcac tggcatcggc tgtttgttct
     2221 tcagcatatt agtaaagcgc tcatttagag acatcttggt ggtgcttttt agcacaactt
     2281 tcggcgctga ctgtgcagcg gtgccaactt ttttgtacaa acttgtcttg tcatcgtcat
     2341 ccttgtaatc gatgtcatga tcatatgtat caccgtcatg gtctttgtag tccatggtgg
     2401 aagcttaagt ttaaacgcta gaggatccag cctccgcgga ggctggatcg gtcccggtgt
     2461 cttctatgga ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa
     2521 cgagctctgc ttatataggg ggtaccttct ctatcactga tagggagctc gagacgtgtc
     2581 gacttctcta tcactgatag ggagacgttc tagacgtacc ttctctatca ctgataggga
     2641 gctcgagacg tgtcgacttc tctatcactg atagggagac gtgcggccag cttggatccc
     2701 tcgagacgcg tatatctggc ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc
     2761 agattcttca tgcaattgtc ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg
     2821 cactactcag cgacctccaa cacacaagca gggagcagat actggcttaa ctatgcggca
     2881 tcagagcaga ttgtactgag agtgcaccat aggggatcgg gagatctccc gatccgtcga
     2941 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3001 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3061 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3121 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3181 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3241 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3301 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3361 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3421 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3481 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3541 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3601 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3661 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3721 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3781 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3841 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3901 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3961 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4021 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4081 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4141 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4201 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4261 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4321 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4381 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4441 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4501 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4561 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4621 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4681 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4741 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4801 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4861 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4921 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4981 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5041 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5101 attacgccaa gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga
     5161 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     5221 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga
     5281 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     5341 cgaagcccaa cctttcatag aagg
//
