LOCUS       sequence-421014-        7326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Kdm1a+exon(12nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7326)
  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 7326)
  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 Kdm1a+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..7326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(81..128)
                     /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     260..279
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    412..636
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     625..642
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    684..708
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3283..3307)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3308..3331)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3483..3507)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3529..3547
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3566..3584
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3603..3621
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3640..3658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3857..3876
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3951..4055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4056..4124,4125..4916)
                     /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(4274..4293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5087..5675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5576..5595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5829..5846
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5963..5984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5999..6004,6005..6022,6023..6029)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6037..6053
                     /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     6042..6064
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6061..6077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6061..6077
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6126..6247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6137..6156)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6191..6210
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6377..7326,1..73))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga
       61 gttcaggctt tttggccaag gaagttccta tactttctag agaataggaa cttcggaata
      121 ggaacttcta ggtacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
      181 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
      241 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
      301 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
      361 gcgcgtgggg atacccccta gagccccagc tggttctttc cgcctcagaa gccatagagc
      421 ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc
      481 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat
      541 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg
      601 ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag cgggtttaaa
      661 cgggccctct agactcgagt cagaccactt tgtacaagaa agttgggtat cacatacttg
      721 gggactgctg tgcagggaca cctggtgtgg cctgacgagg caaagtgtac atggctccca
      781 aaaactggtc ggcaatcctt cctgcttctc gaagcccact caacagagca ccatggactg
      841 tagctgggta gttccggatt gtgtgttctc cagcaaagaa gagtcttggg attggctgtg
      901 gggcacctgg aattgagggg ccaggagtga tcggctgagc cattaaatca tagtcatttc
      961 cagaggatcc tgcagccaca taagaatagg agccccgggc ccacggatca gcacgccaac
     1021 gagataccac agtttccttg ggctgtggga ctgcactgct gccaaaaatc cctttgagaa
     1081 tggccaggca ccggccgaca atcacatcat cactaatgtt ctccataatg ccagcagctt
     1141 ctcctgctac cagggccaat agtattggag ctttatatag gttccagaag aggaagagct
     1201 cacccctgct agcagttgta ctgccaacgt gcccaaacaa attgacactt gggtcccaga
     1261 acacacggtc aaagcataac accaccttgt taaggttgcc aaatcccatc ctttggactg
     1321 cagatgtttt ccactcagga agaggtggca caaactgaac agctggtggc tgctgcttca
     1381 acactcccaa aggaagtgta cagagaactg catcacactt ataaataaag gtttgacttg
     1441 tggaacgtgt gttcacagca atcacttcac atcctgaggc tgtgtagcga acctgccgca
     1501 ctgctgtgtt cagtttaatg tccaagcctt cagctaaagc cacaggcaca catgagtagc
     1561 cattccttac tgtcaggtgg cttccagtaa actcaaagtc atcatcctga tcccaatgtt
     1621 taagagagag ggtagagaga ggtgtggcgt tggcaaattc aagatttgca aaatgccagt
     1681 caagtatttg tctgtctctt gatgagaggt atacatcact tgggggattg gcttccaatt
     1741 cttgaagttt ttcttctagc tttccttgtg tttcagctaa ttcatcatat tccttgcaga
     1801 gggcagtcag gtccctgtgc ttgctcttca ccaggaactc ggctgtgata tctctgggcg
     1861 gcttcacttc tgaagcttct ttgtattgct gatggagctc tttaattttc tccttcaaat
     1921 ttaccatctt attaagaagc tctttcaact cctcctgagt tttcactatc ttcttccaat
     1981 gttcaatctg ctcatctttg acatgctttt cttgcagctg aatgacaacc tccaatgcct
     2041 ggccaaggga tacaggttta ttattgagga cgttgaagtc taactggtga ctaaggtaag
     2101 aagtggcttc tagcaaccgg ttaaattctt gttctaccat ttcatctttt tcttttggaa
     2161 ccttgacagt gtcagcttgt ccattggctt cataaagtgg gcatttttgc ttgatcttgg
     2221 ccagttccat atttacttgt ttgctgacga cagccatggg attccctcca agacctgtta
     2281 caaccatggc gccaagatca gctacatagt ttccttttcg aaatgtagca actcgtccac
     2341 ctactcgatc cctggcttcc agaagtgtga catccatccc aaaactctgt agctgtcgag
     2401 ctgctgccaa gccagaaaca cctgaaccta taataatcac ctttcctgtc tttttaattg
     2461 gtaagggttt tatcctcttg tagatgccga agttgataag accatggcgc tctaagtaac
     2521 tgtgaactcg gtggacaagc acagtatcgc tgttgtaagg cgcttccagc tgctggagag
     2581 tggcttcaaa cgtcagctgg atctttgggt tgtccagcca taactgcaat gtgcgattcc
     2641 tgatgaacag aaaaaccttc tgtgtctgct gaggcccact gatgatgtct gggaaacagg
     2701 ctgcttcctg agaggtcatt cggtcatggg gaagtcggct ttgaaaagct gcaccctcca
     2761 caccagacgg ctcttccggc tcactttcat tttcttcctc aggtggggct tgaggaggtg
     2821 gtgggggaag cttcttttcc ttctctgctt tagcatttct ttcttcttcc gaataatatt
     2881 catcttctga gaggttggcc aagctttcat ccatttctct gtattctacc ttggcccgct
     2941 tgcgccggct ggtccgtcgg ccctccgggg tctcggctat tccggtctcc atgggcgtcg
     3001 cggagccggg cccggctgtg ggccccgcct ggggcccagc agaccccggc ggctcggcta
     3061 ggcccccggg cggcgaggcc cgcggaggct ccttctttcg gggagtgcgc tcgcccgccg
     3121 cccccgtagc catgtcggtg gggcccgtca ggcccgcggg cggcgcggcc acctcagagc
     3181 cgttctcggc accgcccgcc gccccggacc cggcctcggt cccagcagcc gccgccgccg
     3241 ccgctgccgc tgccgccgcc gccgccttct tcccagacaa ggtgccaact tttttgtaca
     3301 aacttgtctt gtcatcgtca tccttgtaat cgatgtcatg atcatatgta tcaccgtcat
     3361 ggtctttgta gtccatggtg gaagcttaag tttaaacgct agaggatcca gcctccgcgg
     3421 aggctggatc ggtcccggtg tcttctatgg aggtcaaaac agcgtggatg gcgtctccag
     3481 gcgatctgac ggttcactaa acgagctctg cttatatagg gggtaccttc tctatcactg
     3541 atagggagct cgagacgtgt cgacttctct atcactgata gggagacgtt ctagacgtac
     3601 cttctctatc actgataggg agctcgagac gtgtcgactt ctctatcact gatagggaga
     3661 cgtgcggcca gcttggatcc ctcgagacgc gtatatctgg cccgtacatc gcgaagcagc
     3721 gcaaaacgcc taaccctaag cagattcttc atgcaattgt cggtcaagcc ttgccttgtt
     3781 gtagcttaaa ttttgctcgc gcactactca gcgacctcca acacacaagc agggagcaga
     3841 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca taggggatcg
     3901 ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3961 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4021 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4081 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4141 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4201 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4261 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4321 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4381 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4441 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4501 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4561 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4621 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4681 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4741 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4801 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4861 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4921 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4981 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5041 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5101 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5161 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5221 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5281 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5341 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5401 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5461 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5521 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5581 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5641 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5701 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5761 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5821 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5881 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5941 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     6001 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     6061 caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga
     6121 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6181 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6241 acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     6301 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     6361 aatctcgtag tacgtgctat tcctttgccc tcggacgagt gctggggcgt cggtttccac
     6421 tatcggcgag tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt
     6481 gtgtacgccc gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc
     6541 aagctgcatc atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc
     6601 ggagcatata cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt
     6661 agcgcgtctg ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct
     6721 cgtattggga atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat
     6781 tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc
     6841 agtcctcggc ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt
     6901 cgtccatcac agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac
     6961 gccatgtagt gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc
     7021 taagatcggc cgcagcgatc gcatccatgg cctccgcgac cggctgcaga acagcgggca
     7081 gttcggtttc aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg
     7141 tcaggctctc gctgaactcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg
     7201 caaagtgccg ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca
     7261 ggacatatcc acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga
     7321 gctgca
//
