LOCUS       sequence-421031-        5679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Sap30bp+exon(48nt)_siResist into N2a 
            FRT rtTA3 expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5679)
  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 5679)
  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 Sap30bp+exon_siResist minimal CMV pC...
FEATURES             Location/Qualifiers
     source          1..5679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    116..237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(153..172)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     207..226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(286..302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(286..302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(299..321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    310..326
                     /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(334..340,341..358,359..364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    379..400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(517..534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(688..1276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(768..787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1447..2238,2239..2307))
                     /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     2070..2089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2308..2412)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2487..2506)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    2705..2723
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2742..2760
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2779..2797
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2816..2834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2856..2880
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3032..3055
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3056..3080
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4008..4032)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4074..4091)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4080..4304
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4437..4456)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4588..4635
                     /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             4643..5665
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
       61 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
      121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      241 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
      301 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      361 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      421 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2401 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     2461 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2521 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     2581 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     2641 gcgctgcttc gcgatgtacg ggccagatat acgcgtctcg agggatccaa gctggccgca
     2701 cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga
     2761 aggtacgtct agaacgtctc cctatcagtg atagagaagt cgacacgtct cgagctccct
     2821 atcagtgata gagaaggtac cccctatata agcagagctc gtttagtgaa ccgtcagatc
     2881 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
     2941 ctccgcggag gctggatcct ctagcgttta aacttaagct tccaccatgg actacaaaga
     3001 ccatgacggt gatacatatg atcatgacat cgattacaag gatgacgatg acaagacaag
     3061 tttgtacaaa aaagttggca ccgcggggaa gaaaaatgtt ttgtcgtctc ttgcaattta
     3121 tgcggaatat tcagaccctg aatctgatgg cgaaactgga gtcgacgctg taggcggagc
     3181 gactgaggag aaaggtggtt tggtatctga cgcctatgga gaagatgact tctctcgccc
     3241 agggggtgat gaggatggtt atgaagaaga ggaggacgag aacagtaagc agtcggaaga
     3301 tgacgattca gagactgaaa aacctgaggc tgatgaccca aaggataaca cagaagcaga
     3361 gaagcgggat ccccaggaat tagtggcctc cttttctgaa agggttcgca acatgtcacc
     3421 cgatgaaatc aagatcccac cagaaccgcc tggcagatgt tctaaccatc tgcaggacaa
     3481 gatccagaag ctctacgagc ggaagataaa ggaggggatg gatatgaact acatcatcca
     3541 gaggaagaag gagttccgca accccagcat ctacgagaaa ctgatccagt tctgtgccat
     3601 tgatgagctg ggcaccaact acccaaagga tatgttcgat ccccacggct ggtctgaaga
     3661 ctcctactat gaggcattag ccaaggccca gaagattgaa atggacaaat tagaaaaggc
     3721 caaaaaggag cgaaccaaaa ttgagtttgt aacgggcacc aagaaaggca ccacgaccaa
     3781 tgccacggcc accagcacca gtactgccag cacagctgtc gcagatgccc agaagaggaa
     3841 gagcaagtgg gactctgcca tccccgtgac gacaatagca cagcccacca tcctcaccac
     3901 cacagccacc ctgccagcag tggtcactgt caccactagt gccagtggct ccaagaccac
     3961 cgtcatctct gctgtgggca ccatcgtgaa gaaggccaag cagtgatacc caactttctt
     4021 gtacaaagtg gtctgactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     4081 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     4141 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4201 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4261 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     4321 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4381 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4441 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4501 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4561 tgattagggt gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga
     4621 aagtatagga acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     4681 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     4741 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     4801 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat
     4861 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg
     4921 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     4981 ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg
     5041 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     5101 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     5161 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     5221 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     5281 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     5341 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     5401 ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     5461 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     5521 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     5581 ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     5641 cactcgtccg agggcaaagg aatagcacgt actacgaga
//
