LOCUS       sequence-421241-        5502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Chtop+exon(138nt)_siResist into N2a FRT
            rtTA3 expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5502)
  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 5502)
  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..5502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    27..148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(64..83)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     118..137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(197..213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(197..213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(210..232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    221..237
                     /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(245..251,252..269,270..275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    290..311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(428..445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(599..1187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(679..698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1358..2149,2150..2218))
                     /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     1981..2000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2219..2323)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2398..2417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    2616..2634
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2653..2671
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2690..2708
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2727..2745
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2767..2791
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2943..2966
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    2967..2991
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3742..3766)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3808..3825)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3814..4038
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4171..4190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4322..4369
                     /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             4377..5399
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
       61 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
      121 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
      181 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
      241 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
      301 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
      421 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      481 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
      541 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      601 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      661 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      721 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      781 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      841 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      901 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      961 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1021 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1081 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1141 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1201 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1261 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1321 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1381 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1441 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1501 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1561 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1621 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1681 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1741 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1801 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1861 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1921 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1981 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2041 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2101 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2161 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2221 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2281 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2341 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     2401 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     2461 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     2521 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     2581 tacgcgtctc gagggatcca agctggccgc acgtctccct atcagtgata gagaagtcga
     2641 cacgtctcga gctccctatc agtgatagag aaggtacgtc tagaacgtct ccctatcagt
     2701 gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta ccccctatat
     2761 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     2821 cctccataga agacaccggg accgatccag cctccgcgga ggctggatcc tctagcgttt
     2881 aaacttaagc ttccaccatg gactacaaag accatgacgg tgatacatat gatcatgaca
     2941 tcgattacaa ggatgacgat gacaagacaa gtttgtacaa aaaagttggc accgctgcac
     3001 agtcagcgcc gaaagttgtg ctaaaaagca ccaccaagat gtctctaaat gagcgcttta
     3061 ctaatatgct gaagaacaaa cagccgatgc cagtgaatat tcgggcttcg atgcagcagc
     3121 agcagcagct agccagtgcc agaaacagaa gactggccca gcagatggag aatagaccct
     3181 ctgtccaggc agcattaaaa cttaagcaga agagcttaaa gcagcgcctg ggtaagagta
     3241 atatccaggc acggttaggc cgacccatag gtgccctggc caggggagca attggaggaa
     3301 gaggcctacc cataatccag agaggcttgc cccgaggagg actacgtggg ggacgtgcta
     3361 ccagaaccct gcttaggggt gggatgtcgc tccgaggtca aaacctgctc cgaggtggac
     3421 gagccgtagc tccccgaatg ggcttaagaa gaggtggtgt tcgaggtcgt ggaggtcctg
     3481 ggagaggggg cctagggcgt ggagctatgg gtcgtggcgg aatcggtggt agaggtcggg
     3541 gtatgatagg tcggggaaga gggggctttg gaggcagagg ccgaggtcgt ggccgaggga
     3601 gaggtgccct cactcgccct gtattgacca aggagcagct ggacaaccaa ttggatgcct
     3661 atatgtccaa gaccaaagga cacctggatg ctgaattgga tgcctacatg gcacagacag
     3721 atcctgaaac caatgattga tacccaactt tcttgtacaa agtggtctga ctcgagtcta
     3781 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3841 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3901 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3961 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     4021 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     4081 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4141 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4201 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4261 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtacct
     4321 agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc ttggccaaaa
     4381 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
     4441 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     4501 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     4561 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt
     4621 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     4681 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
     4741 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     4801 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     4861 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     4921 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     4981 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     5041 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     5101 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     5161 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     5221 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     5281 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     5341 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc
     5401 acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5461 ttttccggga cgccggctgg atgatcctcc agcgcgggga tc
//
