LOCUS       sequence-223646-        7118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7118)
  AUTHORS   Kuo ML, den Besten W, Thomas MC, Sherr CJ
  TITLE     Arf-induced turnover of the nucleolar nucleophosmin-associated 
            SUMO-2/3 protease Senp3.
  JOURNAL   Cell Cycle. 2008 Nov 1;7(21):3378-87. doi: 10.4161/cc.7.21.6930. 
            Epub 2008 Nov 4.
  PUBMED    18948745
REFERENCE   2  (bases 1 to 7118)
  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: 
            "10.4161/cc.7.21.6930"; journalName: "Cell Cycle"; date: "2008-11-1-
            1"; volume: "7"; issue: "21"; pages: "3378-87"
COMMENT     Sequence Label: pCDNA3-FLAG-SENP3
FEATURES             Location/Qualifiers
     source          1..7118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        519..722
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     673..693
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     767..786
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        767..785
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             820..843
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2575..2593)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2576..2593)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2613..2630)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2619..2843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2889..3317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2976..2995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3186..3207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3326..3346)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3331..3660
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3511..3646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3573..3592
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3727..4521
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3781..3800)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4391..4410
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4695..4816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4732..4751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4786..4805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4865..4881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4865..4881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4878..4900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4889..4905
                     /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(4913..4919,4920..4937,4938..4943))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4958..4979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5096..5113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5267..5855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5347..5366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6026..6817,6818..6886))
                     /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     6649..6668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6887..6991)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7066..7085)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
       61 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      121 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      181 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      241 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      481 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      541 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      601 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      661 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      721 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      781 atagggagac ccaagcttgg taccgagctc ggatccatgg attataagga tgacgatgac
      841 aagaaagaga ctatacaggg gaccgggtct tgggggcctg agcctccggg acccggcacc
      901 acttactcaa atcccaggcg agagcgtctt cgttggcccc taccccctaa gccccggctc
      961 aagtccggtg gtggttttgg gccagatcct gggtctggga ccacagtgcc gactagacgc
     1021 ctccctgccc cccggccatc ttttgatgcc tcagctagtg aagaagagga agaggaagag
     1081 gaggaagatg aggaggaagt agcagcttgg aggctacccc ctaggtgggg ccaactgggg
     1141 gcctcccagc gctctcgagc tctccgaccc tctcatagaa aaacctgctc acagcgccgg
     1201 cgccgagcca tgagagcctt ccagatgctg ctctactcaa aaagcacctc gctgacattc
     1261 cactggaagc tttgggggcg ccaccgaggc cggcggcgga acctcgcgca ccccaagaac
     1321 catctctcac cccaggaagg gggtgcaacg ccacaggtgc catcaccctg ctgtcgtttt
     1381 gactcccccc gggggctacc cccaccccgg ctgggtctgc taggtgctct catggcagag
     1441 gatgggatga gagggtctcc accagtgccc tctgggcccc ccatggagga agatggacta
     1501 aggtggaccc caaagtctcc tctggaccct gactcgggcc tcctctcatg tactctaccc
     1561 aatggttttg ggggactgtc tggtcctgaa ggggagcgca gcctggcacc ccctgatgcc
     1621 agcatactca tcagtaatgt gtgcagcatt ggagaccacg tggctcagga actttttcag
     1681 agctctgact tgggcattgc agaagaggca gatcggactg gggagaaagc tggccagcat
     1741 agcccccttc gggaggaaca tgtgacctgt gtgcagagta tcttagatga attccttcaa
     1801 acttatggca gcctcatccc tctcagcact gatgaagttg tagagaagtt ggaggacatt
     1861 ttccagcagg agttctctac accctcaagg aagagtctgg tactacagct gatccagtct
     1921 tatcagcgga tgccaggcaa cgctatggta aggggcttcc gggtatccta taagcgacac
     1981 gtgctcacca tggatgactt gggtacctta tatggacaga actggctcaa tgaccaggtg
     2041 atgaacatgt atggagatct ggtcatggac acagtccctg aaaaggtgca tttcttcaac
     2101 agcttcttct atgataaact ccgtaccaag ggttatgatg gggtaaagag gtggaccaaa
     2161 aatgtggaca tcttcaataa ggaattactg ctaatcccca tccatctgga ggtgcactgg
     2221 tcccttatct cagttgatgt aaggcgacgt accatcacct attttgactc ccagcgaact
     2281 ctaaatcgcc gctgccctaa gcatattgcc aagtatctac aggcagaggc agtcaaaaaa
     2341 gaccgactgg acttccatca gggctggaaa ggttacttca aaatgaatgt ggccaggcag
     2401 aataatgaca gtgactgtgg tgcctttgta ttacagtact gcaagcacct ggccctgtct
     2461 cagccattca gcttcaccca gcaggacatg cccaaactcc gtcgtcagat ctacaaggaa
     2521 ctgtgtcact gcaaactcac tgtgtgactc gagcatgcat ctagagggcc ctattctata
     2581 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     2641 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2701 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2761 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2821 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2881 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2941 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3001 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3061 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3121 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3181 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3241 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3301 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3481 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3661 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3721 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3781 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3841 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3901 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3961 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4021 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4081 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4141 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4201 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4261 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4321 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4381 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4441 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4501 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4561 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4621 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4681 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4741 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4801 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4861 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4921 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4981 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5941 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6001 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6061 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7021 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     7081 tgccgcatag ttaagccagt atctgctccc tgcttgtg
//
