LOCUS       sequence-258817-        6826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses siRNA resistant SENP2 (coiled-coil deletion) 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 6826)
  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 6826)
  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"; 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 CCm
FEATURES             Location/Qualifiers
     source          1..6826
                     /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(2544..2561)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2550..2774
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2907..2926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3058..3105
                     /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             3113..4135
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3113..4135
                     /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    4265..4386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4302..4321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4356..4375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4435..4451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4435..4451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4448..4470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4459..4475
                     /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(4483..4489,4490..4507,4508..4513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4528..4549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4666..4683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4837..5425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4917..4936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5596..6387,6388..6456))
                     /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     6219..6238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6457..6561)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6636..6655)
                     /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 agatcgatta cagatggctg gttaggattc tcggcaccat tttccgtttc tgcgaccggt
      841 cggtgccccc tgcccgggcc ctcctgaaga ggcggcgctc agacagcact ctgttttcta
      901 cagtggacac tgatgaaata ccagccaaaa gaccaagatt agattgcttt attcaccaag
      961 tgaaaaacag tctctacaat gctgccagct tatttggatt cccattccag ctgaccacaa
     1021 agcccatggt aacttctgct tgtaatggaa cacggaatgt ggccccttca ggagaggtat
     1081 tttcgaactc ttcatcttgt gaactgacag gttctggatc ctggaacaac atgctgaaac
     1141 tgggtaataa atctcctaat ggaataagtg actatccaaa gatcagagtg acagttaccc
     1201 gagatcagcc acgcagagtc ctgccttcct ttggttttac tttgaactca gaaggctgta
     1261 atagaagacc aggtggccgt cgccatagca aaggtaatcc agagagttct ttaatgtgga
     1321 aacctcagga acaggctgta acagagatga tttctgaaga gagtggcaag ggtctgaggc
     1381 gtccccattg tggaaactct gtctgtcctg taacttcaaa ttatcacagt tctcaaagaa
     1441 gtcagatgga cacattaaag accaaaggct ggggggaaga gcaaaatcac ggagtcaaaa
     1501 caactcagtt tgttccaaaa caatatagac ttgttgaaac aaggggacct ctatgttcat
     1561 tgagaagtga aaagaggtgt tcaaagggga aaattactga tacagagacg atggtcggaa
     1621 tcagatttga aaatgaaagt aggaggggat accaactgga gcctgaccta tcagaagaag
     1681 tgtcggcccg actccgcctg ggcagtggaa gcaatggctt actcaggagg aaagtgtcaa
     1741 taattgagac aaaggaaaag aattgctcag gcaaagagag ggacagaaga acggacgatc
     1801 tccttgaact tacagaggac atggaaaagg aaatcagtaa tgccctaggc catggcccac
     1861 aggatgaaat cctaagtagt gctttcaaat tgcgaattac tcgaggagac atacaaacat
     1921 taaagaacta tcactggctc aatgatgaag tcattaattt ttacatgaat cttctggtgg
     1981 aaagaaataa aaagcaaggc tatccagcac ttcatgtatt cagtactttc ttctatccta
     2041 aattaaagtc tgggggttac caagcagtga aacgatggac caaaggggta aatctctttg
     2101 aacaagaaat tattctggtg cctattcatc ggaaggtaca ttggagcctg gtggtgattg
     2161 acctaagaaa aaagtgtctt aaatatctgg attctatggg acaaaagggc cacaggatct
     2221 gtgagattct ccttcagtat ttacaggatg aaagtaagac caaaagaaat agtgatctga
     2281 atcttttaga gtggacccat cacagcatga aaccacacga gattcctcaa cagctgaatg
     2341 ggagtgattg tggaatgttt acttgtaaat atgcagatta tatttctagg gacaaaccta
     2401 tcacatttac tcagcaccag atgcctctct tccggaagaa gatggtgtgg gaaatccttc
     2461 atcagcagtt gctgtgaatc gatgatatcc agcacagtgg cggccgctcg agtctagagg
     2521 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2581 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2641 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2701 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2761 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2821 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2881 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2941 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3001 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa
     3061 gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc
     3121 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     3181 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     3241 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     3301 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     3361 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     3421 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     3481 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     3541 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     3601 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     3661 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     3721 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     3781 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     3841 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     3901 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     3961 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     4021 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     4081 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     4141 actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4201 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4261 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4321 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4381 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4441 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4501 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4561 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4621 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4681 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4741 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4801 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4861 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4921 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4981 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5041 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5101 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5161 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5221 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5281 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5341 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5401 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5461 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5521 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5581 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5641 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5701 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5761 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5821 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5881 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5941 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6001 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6061 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6121 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6181 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6241 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6301 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6361 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6421 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6481 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6541 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6601 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6661 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6721 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     6781 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
