LOCUS       sequence-421005-        5679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Sap30bp+exon(48nt) 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 minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..5679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(905..952)
                     /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     1084..1103
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1236..1460
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1449..1466
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    1508..1532
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2460..2484)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2485..2508)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2660..2684)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2706..2724
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2743..2761
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2780..2798
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2817..2835
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3034..3053
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3128..3232
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3233..3301,3302..4093)
                     /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(3451..3470)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4264..4852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4753..4772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5006..5023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5140..5161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5176..5181,5182..5199,5200..5206)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5214..5230
                     /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     5219..5241
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5238..5254
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5238..5254
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5303..5424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5314..5333)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5368..5387
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5554..5679,1..897))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc
       61 gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg
      121 cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca
      181 agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat
      241 cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga
      301 gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag
      361 ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg
      421 atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat
      481 tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc
      541 atccatggcc tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg
      601 caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaactcccc
      661 aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg
      721 atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac
      781 atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct
      841 gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttggc
      901 caaggaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tctaggtacg
      961 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     1021 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     1081 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     1141 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ggggataccc
     1201 cctagagccc cagctggttc tttccgcctc agaagccata gagcccaccg catccccagc
     1261 atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
     1321 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
     1381 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
     1441 ctggcaacta gaaggcacag tcgaggctga tcagcgggtt taaacgggcc ctctagactc
     1501 gagtcagacc actttgtaca agaaagttgg gtatcactgc ttggccttct tcacgatggt
     1561 gcccacagca gagatgacgg tggtcttgga gccactggca ctagtggtga cagtgaccac
     1621 tgctggcagg gtggctgtgg tggtgaggat ggtgggctgt gctattgtcg tcacggggat
     1681 ggcagagtcc cacttgctct tcctcttctg ggcatctgcg acagctgtgc tggcagtact
     1741 ggtgctggtg gccgtggcat tggtcgtggt gcctttcttg gtgcccgtta caaactcaat
     1801 tttggttcgc tcctttttgg ccttttctaa tttgtccatt tcaatcttct gggccttggc
     1861 taatgcctca tagtaggagt cttcagacca gccgtgggga tcgaacatat cctttgggta
     1921 gttggtgccc agctcatcaa tggcacagaa ctggatcagt ttctcgtaga tgctggggtt
     1981 gcggaactcc ttcttcctct ggatgatgta gttcatatcc atcccctcct ttatcttccg
     2041 ctcgtagagc ttctggatct tatcttgtag gtgatttgaa catctgccag gcggttctgg
     2101 tgggatcttg atttcatcgg gtgacatgtt gcgaaccctt tcagaaaagg aggccactaa
     2161 ttcctgggga tcccgcttct ctgcttctgt gttatccttt gggtcatcag cctcaggttt
     2221 ttcagtctct gaatcgtcat cttccgactg cttactgttc tcgtcctcct cttcttcata
     2281 accatcctca tcaccccctg ggcgagagaa gtcatcttct ccataggcgt cagataccaa
     2341 accacctttc tcctcagtcg ctccgcctac agcgtcgact ccagtttcgc catcagattc
     2401 agggtctgaa tattccgcat aaattgcaag agacgacaaa acatttttct tccccgcggt
     2461 gccaactttt ttgtacaaac ttgtcttgtc atcgtcatcc ttgtaatcga tgtcatgatc
     2521 atatgtatca ccgtcatggt ctttgtagtc catggtggaa gcttaagttt aaacgctaga
     2581 ggatccagcc tccgcggagg ctggatcggt cccggtgtct tctatggagg tcaaaacagc
     2641 gtggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt atataggggg
     2701 taccttctct atcactgata gggagctcga gacgtgtcga cttctctatc actgataggg
     2761 agacgttcta gacgtacctt ctctatcact gatagggagc tcgagacgtg tcgacttctc
     2821 tatcactgat agggagacgt gcggccagct tggatccctc gagacgcgta tatctggccc
     2881 gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg
     2941 tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca
     3001 cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag
     3061 tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca cttttcgggg
     3121 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3181 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3241 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3301 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3361 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3421 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3481 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3541 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3601 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3661 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3721 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3781 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3841 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3901 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3961 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4021 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4081 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4141 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4201 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4261 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4321 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4381 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4441 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4501 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4561 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4621 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4681 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4741 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4801 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4861 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4921 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4981 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5041 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5101 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5161 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5221 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga
     5281 ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     5341 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     5401 attataagct gcaataaaca agttggggtg ggcgaagaac tccagcatga gatccccgcg
     5461 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     5521 ggcggcggtg gaatcgaaat ctcgtagtac gtgctattcc tttgccctcg gacgagtgct
     5581 ggggcgtcgg tttccactat cggcgagtac ttctacacag ccatcggtcc agacggccgc
     5641 gcttctgcgg gcgatttgtg tacgcccgac agtcccggc
//
