LOCUS       sequence-258816-        6709 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses siRNA resistant SENP2 (nuclear pore mutant) in mammalian 
            cells, Dox inducible in TetR cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6709)
  AUTHORS   Garvin AJ, Walker AK, Densham RM, Chauhan AS, Stone HR, Mackay HL, 
            Jamshad M, Starowicz K, Daza-Martin M, Ronson GE, Lanz AJ, Beesley 
            JF, Morris JR
  TITLE     The deSUMOylase SENP2 coordinates homologous recombination and 
            nonhomologous end joining by independent mechanisms.
  JOURNAL   Genes Dev. 2019 Mar 1;33(5-6):333-347. doi: 10.1101/gad.321125.118. 
            Epub 2019 Feb 22.
  PUBMED    30796017
REFERENCE   2  (bases 1 to 6709)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes 
            Dev."; date: "2019-03-1"; volume: "33(5-6):333-347. doi"; pages: " 
            10.1101/gad.321125.118. Epub 2019 Feb 22"
COMMENT     Sequence Label: FLAG-SENP2 NPm
FEATURES             Location/Qualifiers
     source          1..6709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             759..782
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2427..2444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2433..2657
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2790..2809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2941..2988
                     /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             2996..4018
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2996..4018
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4148..4269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4185..4204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4239..4258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4318..4334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4318..4334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4331..4353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4342..4358
                     /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(4366..4372,4373..4390,4391..4396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4411..4432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4549..4566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4720..5308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4800..4819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5479..6270,6271..6339))
                     /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     6102..6121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6340..6444)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6519..6538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccatgga ctacaaggac gacgatgaca
      781 agatcgatgc tgccagctta tttggattcc cattccagct gaccacaaag cccatggtaa
      841 cttctgcttg taatggaaca cggaatgtgg ccccttcagg agaggtattt tcgaactctt
      901 catcttgtga actgacaggt tctggatcct ggaacaacat gctgaaactg ggtaataaat
      961 ctcctaatgg aataagtgac tatccaaaga tcagagtgac agttacccga gatcagccac
     1021 gcagagtcct gccttccttt ggttttactt tgaactcaga aggctgtaat agaagaccag
     1081 gtggccgtcg ccatagcaaa ggtaatccag agagttcttt aatgtggaaa cctcaggaac
     1141 aggctgtaac agagatgatt tctgaagaga gtggcaaggg tctgaggcgt ccccattgta
     1201 ctgtggagga gggtgttcaa aaagaggaaa gagagaagta ccgaaagtta ttggaacgac
     1261 ttaaagaaag tggtcatgga aactctgtct gtcctgtaac ttcaaattat cacagttctc
     1321 aaagaagtca gatggacaca ttaaagacca aaggctgggg ggaagagcaa aatcacggag
     1381 tcaaaacaac tcagtttgtt ccaaaacaat atagacttgt tgaaacaagg ggacctctat
     1441 gttcattgag aagtgaaaag aggtgttcaa aggggaaaat tactgataca gagacgatgg
     1501 tcggaatcag atttgaaaat gaaagtagga ggggatacca actggagcct gacctatcag
     1561 aagaagtgtc ggcccgagcc cgcgcgggca gtggaagcaa tggcttactc aggaggaaag
     1621 tgtcaataat tgagacaaag gaaaagaatt gctcaggcaa agagagggac agaagaacgg
     1681 acgatctcct tgaacttaca gaggacatgg aaaaggaaat cagtaatgcc ctaggccatg
     1741 gcccacagga tgaaatccta agtagtgctt tcaaattgcg aattactcga ggagacatac
     1801 aaacattaaa gaactatcac tggctcaatg atgaagtcat taatttttac atgaatcttc
     1861 tggtggaaag aaataaaaag caaggctatc cagcacttca tgtattcagt actttcttct
     1921 atcctaaatt aaagtctggg ggttaccaag cagtgaaacg atggaccaaa ggggtaaatc
     1981 tctttgaaca agaaattatt ctggtgccta ttcatcggaa ggtacattgg agcctggtgg
     2041 tgattgacct aagaaaaaag tgtcttaaat atctggattc tatgggacaa aagggccaca
     2101 ggatctgtga gattctcctt cagtatttac aggatgaaag taagaccaaa agaaatagtg
     2161 atctgaatct tttagagtgg acccatcaca gcatgaaacc acacgagatt cctcaacagc
     2221 tgaatgggag tgattgtgga atgtttactt gtaaatatgc agattatatt tctagggaca
     2281 aacctatcac atttactcag caccagatgc ctctcttccg gaagaagatg gtgtgggaaa
     2341 tccttcatca gcagttgctg tgaaatgata tccagcacag tggcggccgc tcgagtctag
     2401 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2461 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2521 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2581 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2641 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2701 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2761 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2881 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta
     2941 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa
     3001 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     3061 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     3121 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     3181 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     3241 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     3301 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     3361 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     3421 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     3481 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     3541 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     3601 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     3661 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     3721 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     3781 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     3841 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     3901 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     3961 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     4021 cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4081 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4141 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4201 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4261 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4321 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4381 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4441 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4501 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4561 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4621 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4681 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4741 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4801 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4861 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4921 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4981 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5041 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5101 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5161 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5221 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5281 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5341 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5401 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5461 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5521 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5581 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5641 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5701 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5761 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5821 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5881 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5941 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6001 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6061 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6121 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6181 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6241 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6301 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6361 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6421 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     6481 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     6541 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     6601 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     6661 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
