LOCUS       sequence-464793-        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Ubiquitin tagged with a modified Acceptor Peptide (AP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  AUTHORS   Mukhopadhyay U, Levantovsky S, Carusone TM, Gharbi S, Stein F, 
            Behrends C, Bhogaraju S
  TITLE     A ubiquitin-specific, proximity-based labeling approach for the 
            identification of ubiquitin ligase substrates.
  JOURNAL   Sci Adv. 2024 Aug 9;10(32):eadp3000. doi: 10.1126/sciadv.adp3000. 
            Epub 2024 Aug 9.
  PUBMED    39121224
REFERENCE   2  (bases 1 to 5431)
  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: "Sci Adv."; 
            date: "2024-08-9"; volume: "10(32):eadp3000. doi"; pages: " 
            10.1126/sciadv.adp3000. Epub 2024 Aug 9"
COMMENT     Sequence Label: pcDNA5-AP(-3)-Ub
FEATURES             Location/Qualifiers
     source          1..5431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    841..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(878..897)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     932..951
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1011..1027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1011..1027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1024..1046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1035..1051
                     /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(1059..1065,1066..1083,1084..1089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1104..1125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1242..1259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1413..2001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1493..1512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2172..2963,2964..3032))
                     /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     2795..2814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3033..3137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3212..3231)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3403..3782
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3783..3986
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3937..3957
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    3988..4006
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4009..4027
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4037..4061
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4224..4451
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     primer_bind     complement(4551..4568)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4557..4781
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4914..4933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5065..5112
                     /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(5120..5431,1..711)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(5120..5431,1..711)
                     /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
                     "
ORIGIN
        1 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
       61 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
      121 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
      181 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
      241 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
      301 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
      361 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
      421 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
      481 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
      541 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
      601 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
      661 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtacta
      721 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
      781 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
      841 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
      901 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      961 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     1021 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     3181 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3241 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     3301 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     3361 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     3421 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     3481 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     3541 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     3601 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     3661 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     3721 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     3781 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     3841 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     3901 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     3961 ggtgggaggt ctatataagc agagctctcc ctatcagtga tagagatctc cctatcagtg
     4021 atagagatcg tcgacgagct cgtttagtga accgtcagat cgcctggaga cgccatccac
     4081 gctgttttga cctccataga agacaccggg accgatccag cctccggact ctagcgttta
     4141 aacttaagat gggcctgaac gacatcttcg aggcccagaa gatcgagggg gagttcggct
     4201 ccggctccac ctccggctcc ggcatgcaga tcttcgtgaa gactctgact ggtaagacca
     4261 tcaccctcga ggttgagccc agtgacacca ttgagaatgt caaggcaaag atccaagata
     4321 aggaaggcat ccctcctgac cagcagaggc tgatctttgc tggaaaacag ctggaagatg
     4381 ggcgcaccct gtctgactac aacatccaga aagagtccac cctgcacctg gtactccgtc
     4441 tcagaggtgg gtaataagga tccactagtc cagtgtggtg gaattctgca gatatccagc
     4501 acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     4561 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4621 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4681 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4741 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4801 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4861 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4921 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4981 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5041 ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag
     5101 tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     5161 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     5221 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     5281 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     5341 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc
     5401 acagggtgtc acgttgcaag acctgcctga a
//
