LOCUS       sequence-421017-        7464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Kansl3+exon(78nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7464)
  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 7464)
  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 Kansl3+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..7464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1008..1129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1045..1064)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1099..1118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1178..1194)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1178..1194)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1191..1213)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1202..1218
                     /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(1226..1232,1233..1250,1251..1256))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1271..1292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1409..1426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1580..2168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1660..1679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2339..3130,3131..3199))
                     /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     2962..2981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3200..3304)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3379..3398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3597..3615
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3634..3652
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3671..3689
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3708..3726
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3748..3772
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3924..3947
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3948..3972
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6013..6024
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(6685..6709)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6751..6768)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6757..6981
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7114..7133)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    7265..7312
                     /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(7320..7464,1..878)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg
       61 ccgcgctccc gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt
      121 gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg
      181 ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga
      241 cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt
      301 tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
      361 tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg
      421 aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc
      481 gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg
      541 ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg
      601 agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg
      661 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc
      721 agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg
      781 cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa
      841 accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga gatttcgatt
      901 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
      961 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     1021 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1081 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     1141 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1201 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1261 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1321 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1381 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1441 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1501 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1561 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1621 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1681 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1741 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1801 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1861 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1921 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1981 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2041 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2101 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2161 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2221 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2281 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2401 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2521 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2641 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2761 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2941 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3001 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3301 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     3361 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     3421 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     3481 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     3541 tcgcgatgta cgggccagat atacgcgtct cgagggatcc aagctggccg cacgtctccc
     3601 tatcagtgat agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtacgt
     3661 ctagaacgtc tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga
     3721 tagagaaggt accccctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     3781 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccgcgg
     3841 aggctggatc ctctagcgtt taaacttaag cttccaccat ggactacaaa gaccatgacg
     3901 gtgatacata tgatcatgac atcgattaca aggatgacga tgacaagaca agtttgtaca
     3961 aaaaagttgg caccgcccac cggggtgggg agagggactt ccagacttca gctcggcgga
     4021 tgggtacctc cctgctcttc cagctttcag tgcatgagcg ggaactggac ctcgttttcc
     4081 tggatcatag ctatgcaaaa ccgtggagtg cccacccaga tgccagtagt gcccgcccca
     4141 cccgcatgct ctttgtaacc ccccgtcggc agcaggagaa tactattgag tcagacgttc
     4201 caatagatgt ggagacggtc acggcaactc ctgttccact ttatgacaat cagaaggcaa
     4261 gaagtgtgat gaacgagtgt gaacggcacg tgatcttcgc caggacagat gcagatgctc
     4321 ctcctccgcc agaagactgg gaagaacatg tcaacaggac tggctggaca gtggctcaga
     4381 ataagctatt taacaagatc ctcaaagccc tgcagtctga ccggcttgca cgattggcca
     4441 atgaaggggc ttgcaatgag cctgtgctgc gtcgcgtggc tgtggacaag tgtgcaagga
     4501 gggtgcggca ggcgctggca agtgtcagct gggacaccaa gctgacccag tggctgcaca
     4561 ccaccctcgt ggagaccctg agcctgccca tgctggctgc ttacttggat gccctgcaga
     4621 cactgaaagg gaagatccca actttgatag atcggatgct tgtgtcgtcc aacaccaaaa
     4681 ctggggctgc aggagcggag gccttgtccc ttctcctcaa gaggccctgg gaccctgctg
     4741 ttggtgtgct ttcacataac aaaccaagca agctccctgg ttctcctttg attctcatcg
     4801 tctcctctgg tccctccagc tctgtgttcc ctgcctcacg ccgccaccgc ttctggcagt
     4861 ctcagctctc ctgcttaggc aaggtcatcc ctgtagcgac ccatctactg aacaatggca
     4921 gtggggtagg agttctgcag tgcctcgaac acatgattgg ggccgtgaga agcaaagtgc
     4981 tagagattca cagccatttc ccacacaaac ccatcatcct gattggatgg aatacgggag
     5041 ctttggtggc ctgtcatgtg tctgtgatgg agtacgtcac tgctgttgtc tgccttgggt
     5101 ttcctctgct tactgtggat ggacccaggg gggatgtgga cgatcccctc ctggacatga
     5161 agactccagt cctcttcgtc attggtcaga attctctaca gtgtcaccct gaagccatgg
     5221 aggacttccg ggaaaagatt cgagcagaaa atagcttggt ggtggttggg ggagctgatg
     5281 ataatctcag aataagcaaa gcaaagaaga aatcagaggg cctgactcag agcatggtgg
     5341 acagatgtat tcaggatgag attgtagact tcctgactgg agtgctcact cgtgctgagg
     5401 gtcatgtggg ctctgagcct cgggatcagg atgctgagaa gaagaagaaa ccccgggact
     5461 tgacccgaag agacctggcc tttgaaattc ctgagcgagg cagtcgtcct gcttccccag
     5521 ctgccaggct ccccacatcc ccctcaggct cagaggatct ctccagtgtg tccagcagcc
     5581 ccacctctag ccctaagacc aaagtgacca cagtgacctc tacccagaaa tcaagtcaga
     5641 ttgggacctc ccagctgctg aaaagacatg tgcagaggac agaagctgtg ctgacccaca
     5701 gacaagccca agcacagttt gctgcttttc tgaaacaaaa catgctggtg aggaaagctt
     5761 ttcctcctgg cacctcctcc tgtctctttg ttcccatttc atcagaatca gtggaggaca
     5821 tagagaaaga agagcttcgg gtccagctta agaggcacca ttcttccagt cctcttcctg
     5881 gagcaaagcc ttccaagcga ccaaagatca aagtgtccct gatctcccaa ggggacacag
     5941 ttggaggacc ttgcactctt tctcaaggtg gaacacctga agctgccgga gggaagccca
     6001 tcaccatgac cctgggagct tcagcaggag ccaaggaact cactggactt ctcactacag
     6061 ccaagtcaag ttcttctgaa ggtggaggta cagccagcac aaccccatct gtggcttcca
     6121 gcagtgccac acctaatgct atccacacgc tgcagagccg cttggtggcc acttctccag
     6181 gcagctccct tccagggacc gcttcagcca gcagcctcct ccagggcctc agcttcagct
     6241 tacaggatat tagcagcaag acctctggcc tcccaggaag tccctcccca gggccagccc
     6301 cacaggccac cagtgtgaaa ttgccaaccc ccatgcagag tctgggtgcc atcaccacag
     6361 gcacgagcac cattgtccgt accatccctg tggccaccac tctctcctcc ttgggtgcca
     6421 ctcctggtgg gaagcccact gccatccacc agctgctgac caatgggagc ctcgctaagt
     6481 tggcaagcag cctacctggc ttggctcaga tttccaacca agcatcaggc ttaaaggtcc
     6541 ccaccaccat tactctgacg ctgcgtggcc agccaagcag aattaccacg ctaagcccta
     6601 tggggtcagg ggcaacgccg tccgaggagc ccaactccca gatgctacca tctagttcac
     6661 agcgcctgcc tccagcaccc tgatacccaa ctttcttgta caaagtggtc tgactcgagt
     6721 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     6781 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     6841 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     6901 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     6961 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     7021 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     7081 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     7141 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     7201 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     7261 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca
     7321 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     7381 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     7441 gagggcgtgg atatgtcctg cggg
//
