LOCUS       sequence-421027-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Sap30bp-exon_siResist into N2a FRT 
            rtTA3 expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  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 5631)
  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 minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    92..213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(129..148)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     183..202
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(262..278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(262..278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(275..297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    286..302
                     /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(310..316,317..334,335..340))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    355..376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(493..510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(664..1252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(744..763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1423..2214,2215..2283))
                     /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     2046..2065
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2284..2388)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2463..2482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    2681..2699
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2718..2736
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2755..2773
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2792..2810
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2832..2856
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3008..3031
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3032..3056
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3936..3960)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4002..4019)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4008..4232
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4365..4384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4516..4563
                     /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             4571..5593
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
       61 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
      121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
      181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
      241 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
      301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
      361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
      481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1261 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1321 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1381 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1441 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1501 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1561 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1621 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1681 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1741 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1801 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1861 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1921 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1981 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2041 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2101 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2161 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2221 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2281 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2341 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2401 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     2461 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     2521 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     2581 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     2641 agatatacgc gtctcgaggg atccaagctg gccgcacgtc tccctatcag tgatagagaa
     2701 gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt acgtctagaa cgtctcccta
     2761 tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga aggtaccccc
     2821 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
     2881 tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggaggctg gatcctctag
     2941 cgtttaaact taagcttcca ccatggacta caaagaccat gacggtgata catatgatca
     3001 tgacatcgat tacaaggatg acgatgacaa gacaagtttg tacaaaaaag ttggcaccgc
     3061 ggggaagaaa aatgttttgt cgtctcttgc aatttatgcg gaatattcag accctgaatc
     3121 tgatggcgaa actggagtcg acgctgtagg cggagcgact gaggagaaag gtggtttggt
     3181 atctgacgcc tatggagaag atgacttctc tcgcccaggg ggtgatgagg atggttatga
     3241 agaagaggag gacgagaaca gtaagcagtc ggataacaca gaagcagaga agcgggatcc
     3301 ccaggaatta gtggcctcct tttctgaaag ggttcgcaac atgtcacccg atgaaatcaa
     3361 gatcccacca gaaccgcctg gcagatgttc taaccatctg caggacaaga tccagaagct
     3421 ctacgagcgg aagataaagg aggggatgga tatgaactac atcatccaga ggaagaagga
     3481 gttccgcaac cccagcatct acgagaaact gatccagttc tgtgccattg atgagctggg
     3541 caccaactac ccaaaggata tgttcgatcc ccacggctgg tctgaagact cctactatga
     3601 ggcattagcc aaggcccaga agattgaaat ggacaaatta gaaaaggcca aaaaggagcg
     3661 aaccaaaatt gagtttgtaa cgggcaccaa gaaaggcacc acgaccaatg ccacggccac
     3721 cagcaccagt actgccagca cagctgtcgc agatgcccag aagaggaaga gcaagtggga
     3781 ctctgccatc cccgtgacga caatagcaca gcccaccatc ctcaccacca cagccaccct
     3841 gccagcagtg gtcactgtca ccactagtgc cagtggctcc aagaccaccg tcatctctgc
     3901 tgtgggcacc atcgtgaaga aggccaagca gtgataccca actttcttgt acaaagtggt
     3961 ctgactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     4021 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4081 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4141 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4201 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4261 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4321 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4381 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4441 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4501 tggttcacgt acctagaagt tcctattccg aagttcctat tctctagaaa gtataggaac
     4561 ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4621 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4681 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4741 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     4801 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     4861 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     4921 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4981 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     5041 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     5101 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     5161 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     5221 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     5281 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     5341 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     5401 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     5461 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     5521 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     5581 ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct a
//
