LOCUS       sequence-420974-        6270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Mllt1-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6270)
  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 6270)
  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..6270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(609..656)
                     /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     788..807
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    940..1164
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1153..1170
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    1212..1236
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    1664..1675
                     /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(2755..2779)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2780..2803)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2955..2979)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3001..3019
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3038..3056
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3075..3093
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3112..3130
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3329..3348
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3423..3527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3528..3596,3597..4388)
                     /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(3746..3765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4559..5147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5048..5067
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5301..5318
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5435..5456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5471..5476,5477..5494,5495..5501)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5509..5525
                     /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     5514..5536
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5533..5549
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5533..5549
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5598..5719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5609..5628)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5663..5682
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5849..6270,1..601))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca
       61 tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc
      121 agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac
      181 cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga
      241 tcgcatccat ggcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt
      301 cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaact
      361 ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat
      421 aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc
      481 ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga
      541 cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt
      601 tggccaagga agttcctata ctttctagag aataggaact tcggaatagg aacttctagg
      661 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
      721 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
      781 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
      841 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat
      901 accccctaga gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc
      961 cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
     1021 ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa
     1081 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
     1141 atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctctag
     1201 actcgagtca gaccactttg tacaagaaag ttgggtatca tgtggccacg gcctccaggc
     1261 agctctgcag cttgcgaaca gtgctttcat ccaaggagaa aaggtcaaag tcgaaggtgg
     1321 tgttggtgac attgaagtgg ccagtctcct cgatcagatt gacaatctgc tgcagcacat
     1381 tgcgctcgcg cagagccatc agtctccggt gcagctccac cagctcatcc gtgtaagcct
     1441 tgtcataggt agccttcttc agcatctttt cgggcttgct gcagggctcg gggctcctcc
     1501 gaccagacac cttgctgctg ggagggggcg gcttctgggg aggcaggggc tcacggccag
     1561 gcaggcagga gtctgcgctg ttgtcactct cgctgtcact gaagcttagc ctggactccc
     1621 ggcctgggtt ggccttgccg gtgttctcct ctcctgacga tgagtcatcc tcatcagagc
     1681 cctcagactg caggtcctcc accatggagc gcaggggtcc ttggctgtgg ttctgagacg
     1741 gctcaaagtc tgagtcagag ctggagtcag agctggagct ggcattagct gggcttgact
     1801 gggccgactc tgaccggaag gaggcctcat cctctgagtt tgagtcttca gtctgcagag
     1861 tccgcttggc ctcccgaggc tcactctctg ccttcccctt gtcacctttg ctgtttttgt
     1921 ccctggccac cttcttgtca ggaaaggagg aagctcgggg tgaggtgcct ggtgcacgcc
     1981 gtgtcccctt tgacccactc tttttaggct ttttggcgct gagcttcggt gagtccgtgg
     2041 cagctggcct cttgctggag gccttgggca gggtaggagg ctggggcacc cccttggggg
     2101 acaggctctc cagcttggtt tcctttaggg ccatcttggg ctccttgaag gcagccttgg
     2161 gcactggggc cttctcctcc ttgggcagct tgcgtgcagc atccttggca ctcttgggct
     2221 cacgctccgg ctccttggag caactcctgc tctctgagtc cttgcgcggc cgctcacggt
     2281 gctccctggc caccttgtgt ggtttggaag ccttgccact ctccttgttg gcatcccctc
     2341 cagccatgag cagcttgcag cgaaactcgg tggtggggtt attgaaggtg agcttctcac
     2401 agcgcaggtg gttgacagga gggttgccct ccaggttgag aaacaggtcg taggtgaagc
     2461 agaccttcct cggctcctcc ttgtttttga agtacacttc aatgagcatg atgaagcctg
     2521 catagcccga ctcctccact ttgtaggggg gctccttgca cacacgcttg ggtttaggga
     2581 agctgtcatg cagccggaag atgaccttct ccacaaagtg ctggatgtca cactgctcag
     2641 ggccacgcac gaagaccatc cagtcatggg tgaagccctc ggtggtgggc ttcttccgta
     2701 gctgggcacg gtgccccagc tctaacttca cctggacagt gcactgattg tcggtgccaa
     2761 cttttttgta caaacttgtc ttgtcatcgt catccttgta atcgatgtca tgatcatatg
     2821 tatcaccgtc atggtctttg tagtccatgg tggaagctta agtttaaacg ctagaggatc
     2881 cagcctccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga
     2941 tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata gggggtacct
     3001 tctctatcac tgatagggag ctcgagacgt gtcgacttct ctatcactga tagggagacg
     3061 ttctagacgt accttctcta tcactgatag ggagctcgag acgtgtcgac ttctctatca
     3121 ctgataggga gacgtgcggc cagcttggat ccctcgagac gcgtatatct ggcccgtaca
     3181 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag
     3241 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa
     3301 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3361 cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
     3421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3541 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3601 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3661 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3721 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3781 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3841 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3901 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3961 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4021 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4081 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4141 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4201 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4261 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4321 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4381 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4441 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4501 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4561 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4621 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4681 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4741 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4801 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4861 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4921 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4981 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5041 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5101 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5161 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5221 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5281 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5341 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     5401 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     5461 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     5521 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     5581 acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     5641 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     5701 aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga
     5761 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     5821 cggtggaatc gaaatctcgt agtacgtgct attcctttgc cctcggacga gtgctggggc
     5881 gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc
     5941 tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc
     6001 gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt
     6061 caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc
     6121 tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga
     6181 tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg
     6241 ttatgcggcc attgtccgtc aggacattgt
//
