LOCUS       sequence-421021-        7386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Kansl3-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7386)
  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 7386)
  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..7386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    969..1090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1006..1025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1060..1079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1139..1155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1139..1155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1152..1174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1163..1179
                     /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(1187..1193,1194..1211,1212..1217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1232..1253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1370..1387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1541..2129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1621..1640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2300..3091,3092..3160))
                     /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     2923..2942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3161..3265)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3340..3359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3558..3576
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3595..3613
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3632..3650
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3669..3687
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3709..3733
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3885..3908
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3909..3933
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5896..5907
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(6568..6592)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6634..6651)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6640..6864
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6997..7016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    7148..7195
                     /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(7203..7386,1..839)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt
       61 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
      121 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
      181 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
      241 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
      301 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
      361 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
      421 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
      481 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
      541 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
      601 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
      661 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
      721 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
      781 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc
      841 acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
      901 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
      961 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1021 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1081 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     1141 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1201 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1261 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1321 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1381 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1441 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1501 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1561 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1621 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1681 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1741 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1801 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1861 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1921 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1981 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2041 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2101 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2161 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2221 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2281 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2341 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2401 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2461 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2521 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2581 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2641 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2701 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2761 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2821 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2881 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2941 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3001 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3061 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3121 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3181 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3241 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     3301 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     3361 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     3421 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     3481 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtc tcgagggatc
     3541 caagctggcc gcacgtctcc ctatcagtga tagagaagtc gacacgtctc gagctcccta
     3601 tcagtgatag agaaggtacg tctagaacgt ctccctatca gtgatagaga agtcgacacg
     3661 tctcgagctc cctatcagtg atagagaagg taccccctat ataagcagag ctcgtttagt
     3721 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
     3781 ggaccgatcc agcctccgcg gaggctggat cctctagcgt ttaaacttaa gcttccacca
     3841 tggactacaa agaccatgac ggtgatacat atgatcatga catcgattac aaggatgacg
     3901 atgacaagac aagtttgtac aaaaaagttg gcaccgccca ccggggtggg gagagggact
     3961 tccagacttc agctcggcgg atgggtacct ccctgctctt ccagctttca gtgcatgagc
     4021 gggaactgga cctcgttttc ctggatcata gctatgcaaa accgtggagt gcccacccag
     4081 atgccagtag tgcccgcccc acccgcatgc tctttgtaac cccccgtcgg cagcaggaga
     4141 atactattga gtcagacgtt ccaatagatg tggagacggt cacggcaact cctgttccac
     4201 tttatgacaa tcagaaggca agaagtgtga tgaacgagtg tgaacggcac gtgatcttcg
     4261 ccaggacaga tgcagatgct cctcctccgc cagaagactg ggaagaacat gtcaacagga
     4321 ctggctggac agtggctcag aataagctat ttaacaagat cctcaaagcc ctgcagtctg
     4381 accggcttgc acgattggcc aatgaagggg cttgcaatga gcctgtgctg cgtcgcgtgg
     4441 ctgtggacaa gtgtgcaagg agggtgcggc aggcgctggc aagtgtcagc tgggacacca
     4501 agctgaccca gtggctgcac accaccctcg tggagaccct gagcctgccc atgctggctg
     4561 cttacttgga tgccctgcag acactgaaag ggaagatccc aactttgata gatcggatgc
     4621 ttgtgtcgtc caacaccaaa actggggctg caggagcgga ggccttgtcc cttctcctca
     4681 agaggccctg ggaccctgct gttggtgtgc tttcacataa caaaccaagc aagctccctg
     4741 gttctccttt gattctcatc gtctcctctg gtccctccag ctctgtgttc cctgcctcac
     4801 gccgccaccg cttctggcag tctcagctct cctgcttagg caaggtcatc cctgtagcga
     4861 cccatctact gaacaatggc agtggggtag gagttctgca gtgcctcgaa cacatgattg
     4921 gggccgtgag aagcaaagtg ctagagattc acagccattt cccacacaaa cccatcatcc
     4981 tgattggatg gaatacggga gctttggtgg cctgtcatgt gtctgtgatg gagtacgtca
     5041 ctgctgttgt ctgccttggg tttcctctgc ttactgtgga tggacccagg ggggatgtgg
     5101 acgatcccct cctggacatg aagactccag tcctcttcgt cattggtcag aattctctac
     5161 agtgtcaccc tgaagccatg gaggacttcc gggaaaagat tcgagcagaa aatagcttgg
     5221 tggtggttgg gggagctgat gataatctca gaataagcaa agcaaagaag aaatcagagg
     5281 gcctgactca gagcatggtg gacagatgta ttcaggatga gattgtagac ttcctgactg
     5341 gagtgctcac tcgtgctgag ggtcatgtgg gctctgagcc tcgggatcag gatgctgaga
     5401 agaagaagaa accccgggac ttgacccgaa gagacctggc ctttgaaatt cctgagcgag
     5461 gcagtcgtcc tgcttcccca gctgccaggc tccccacatc cccctcaggc tcagaggatc
     5521 tctccagtgt gtccagcagc cccacctcta gccctaagac caaagtgacc acagtgacct
     5581 ctacccagaa atcaagtcag attgggacct cccagctgct gaaaagacat gtgcagagga
     5641 cagaagctgt gctgacccac agacaagccc aagttcccat ttcatcagaa tcagtggagg
     5701 acatagagaa agaagagctt cgggtccagc ttaagaggca ccattcttcc agtcctcttc
     5761 ctggagcaaa gccttccaag cgaccaaaga tcaaagtgtc cctgatctcc caaggggaca
     5821 cagttggagg accttgcact ctttctcaag gtggaacacc tgaagctgcc ggagggaagc
     5881 ccatcaccat gaccctggga gcttcagcag gagccaagga actcactgga cttctcacta
     5941 cagccaagtc aagttcttct gaaggtggag gtacagccag cacaacccca tctgtggctt
     6001 ccagcagtgc cacacctaat gctatccaca cgctgcagag ccgcttggtg gccacttctc
     6061 caggcagctc ccttccaggg accgcttcag ccagcagcct cctccagggc ctcagcttca
     6121 gcttacagga tattagcagc aagacctctg gcctcccagg aagtccctcc ccagggccag
     6181 ccccacaggc caccagtgtg aaattgccaa cccccatgca gagtctgggt gccatcacca
     6241 caggcacgag caccattgtc cgtaccatcc ctgtggccac cactctctcc tccttgggtg
     6301 ccactcctgg tgggaagccc actgccatcc accagctgct gaccaatggg agcctcgcta
     6361 agttggcaag cagcctacct ggcttggctc agatttccaa ccaagcatca ggcttaaagg
     6421 tccccaccac cattactctg acgctgcgtg gccagccaag cagaattacc acgctaagcc
     6481 ctatggggtc aggggcaacg ccgtccgagg agcccaactc ccagatgcta ccatctagtt
     6541 cacagcgcct gcctccagca ccctgatacc caactttctt gtacaaagtg gtctgactcg
     6601 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     6661 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     6721 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     6781 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     6841 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     6901 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6961 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     7021 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     7081 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     7141 gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg
     7201 ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     7261 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     7321 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     7381 gttatg
//
