LOCUS       sequence-420982-        6396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Mllt1+exon(126nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6396)
  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 6396)
  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 Mllt1+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    474..595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(511..530)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     565..584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(644..660)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(644..660)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(657..679)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    668..684
                     /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(692..698,699..716,717..722))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    737..758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(875..892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1046..1634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1126..1145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1805..2596,2597..2665))
                     /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     2428..2447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2666..2770)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2845..2864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3063..3081
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3100..3118
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3137..3155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3174..3192
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3214..3238
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3390..3413
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3414..3438
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4644..4655
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    complement(5083..5107)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5149..5166)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5155..5379
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5512..5531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5663..5710
                     /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             join(5718..6396,1..344)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
       61 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
      121 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
      181 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
      241 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
      301 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
      361 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
      421 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
      481 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
      541 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
      601 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
      661 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      721 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      781 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      841 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      901 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      961 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1021 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1081 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1141 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1201 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1261 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1321 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1381 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1441 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1501 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1561 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1621 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1681 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1741 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1801 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1861 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1921 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1981 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2041 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2101 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2161 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2221 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2281 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2341 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2401 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2461 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2521 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2581 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2641 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2701 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2761 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     2821 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2881 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     2941 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     3001 gctgcttcgc gatgtacggg ccagatatac gcgtctcgag ggatccaagc tggccgcacg
     3061 tctccctatc agtgatagag aagtcgacac gtctcgagct ccctatcagt gatagagaag
     3121 gtacgtctag aacgtctccc tatcagtgat agagaagtcg acacgtctcg agctccctat
     3181 cagtgataga gaaggtaccc cctatataag cagagctcgt ttagtgaacc gtcagatcgc
     3241 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     3301 ccgcggaggc tggatcctct agcgtttaaa cttaagcttc caccatggac tacaaagacc
     3361 atgacggtga tacatatgat catgacatcg attacaagga tgacgatgac aagacaagtt
     3421 tgtacaaaaa agttggcacc gacaatcagt gcactgtcca ggtgaagtta gagctggggc
     3481 accgtgccca gctacggaag aagcccacca ccgagggctt cacccatgac tggatggtct
     3541 tcgtgcgtgg ccctgagcag tgtgacatcc agcactttgt ggagaaggtc atcttccggc
     3601 tgcatgacag cttccctaaa cccaagcgtg tgtgcaagga gcccccctac aaagtggagg
     3661 agtcgggcta tgcaggcttc atcatgctca ttgaagtgta cttcaaaaac aaggaggagc
     3721 cgaggaaggt ctgcttcacc tacgacctgt ttctcaacct ggagggcaac cctcctgtca
     3781 accacctgcg ctgtgagaag ctcaccttca ataaccccac caccgagttt cgctgcaagc
     3841 tgctcatggc tggaggggtg atggtaatgc ccgaaggagc ggacacggtg tccaggccca
     3901 gccctgacta ccccatgtta cccacaattc cactctctgc cttctctgac cccaagaaga
     3961 acaaaccatc ccacggctcc aaggatgcca acaaggagag tggcaaggct tccaaaccac
     4021 acaaggtggc cagggagcac cgtgagcggc cgcgcaagga ctcagagagc aggagttgct
     4081 ccaaggagcc ggagcgtgag cccaagagtg ccaaggatgc tgcacgcaag ctgcccaagg
     4141 aggagaaggc cccagtgccc aaggctgcct tcaaggagcc caagatggcc ctaaaggaaa
     4201 ccaagctgga gagcctgtcc cccaaggggg tgccccagcc tcctaccctg cccaaggcct
     4261 ccagcaagag gccagctgcc acggactcac cgaagctcag cgccaaaaag cctaaaaaga
     4321 gtgggtcaaa ggggacacgg cgtgcaccag gcacctcacc ccgagcttcc tcctttcctg
     4381 acaagaaggt ggccagggac aaaaacagca aaggtgacaa ggggaaggca gagagtgagc
     4441 ctcgggaggc caagcggact ctgcagactg aagactcaaa ctcagaggat gaggcctcct
     4501 tccggtcaga gtcggcccag tcaagcccag ctaatgccag ctccagctct gactccagct
     4561 ctgactcaga ctttgagccg tctcagaacc acagccaagg acccctgcgc tccatggtgg
     4621 aggacctgca gtctgagggc tctgatgagg atgactcatc gtcaggagag gagaacaccg
     4681 gcaaggccaa cccaggccgg gagtccaggc taagcttcag tgacagcgag agtgacaaca
     4741 gcgcagactc ctgcctgcct ggccgtgagc ccctgcctcc ccagaagccg ccccctccca
     4801 gcagcaaggt gtctggtcgg aggagccccg agccctgcag caagcccgaa aagatgctga
     4861 agaaggctac ctatgacaag gcttacacgg atgagctggt ggagctgcac cggagactga
     4921 tggctctgcg cgagcgcaat gtgctgcagc agattgtcaa tctgatcgag gagactggcc
     4981 acttcaatgt caccaacacc accttcgact ttgacctttt ctccttggat gaaagcactg
     5041 ttcgcaagct gcagagctgc ctggaggccg tggccacatg atacccaact ttcttgtaca
     5101 aagtggtctg actcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     5161 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     5221 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     5281 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     5341 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     5401 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5461 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5521 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5581 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5641 agggtgatgg ttcacgtacc tagaagttcc tattccgaag ttcctattct ctagaaagta
     5701 taggaacttc cttggccaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
     5761 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     5821 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     5881 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     5941 aagtgcttga cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     6001 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
     6061 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     6121 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     6181 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     6241 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     6301 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     6361 actggagcga ggcgatgttc ggggattccc aatacg
//
