LOCUS       sequence-313931-        7822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7822)
  AUTHORS   Sowa ME, Bennett EJ, Gygi SP, Harper JW
  TITLE     Defining the human deubiquitinating enzyme interaction landscape.
  JOURNAL   Cell. 2009 Jul 23. 138(2):389-403.
  PUBMED    19615732
REFERENCE   2  (bases 1 to 7822)
  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: "Cell. 2009 
            Jul 23. 138(2):389-403."
COMMENT     Sequence Label: Flag-HA-USP45
FEATURES             Location/Qualifiers
     source          1..7822
                     /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"
     regulatory      732..741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             741..764
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             777..803
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    819..843
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1726..1750)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        1769..2268
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1804..1827)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2201..2220
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2289..2888
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2289..2308)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2785..2805
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    3474..3490
                     /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(3498..3504,3505..3522,3523..3528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3543..3564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3681..3698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3852..4440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3932..3951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4611..5402,5403..5471))
                     /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     5234..5253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5472..5576)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5644..5662
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5700..5722)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5822..5841
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6035..6057
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6185..6204
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6404..6920
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6984..7325
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7392..7808
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7738..7760
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7776..7792
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
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 tccggggatc tgccaccatg gactacaagg atgacgatga caagctcgat ggaggatacc
      781 catacgatgt tccagattac gctgctcgag gttatcaaac aagtttgtac aaaaaagttg
      841 gcgcgcccac catgcgggtg aaagatccaa ctaaagcttt acctgagaaa gccaaaagaa
      901 gtaaaaggcc tactgtacct catgatgaag actcttcaga tgatattgct gtaggtttaa
      961 cttgccaaca tgtaagtcat gctatcagcg tgaatcatgt aaagagagca atagctgaga
     1021 atctgtggtc agtttgctca gaatgtttag aagaaagaag attctatgat gggcagctag
     1081 tacttacttc tgatatttgg ttgtgcctca agtgtggctt ccagggatgt ggtaaaaact
     1141 cagaaagcca acattcattg aagcacttta agagttccag aacagagccc cattgtatta
     1201 taattaatct gagcacatgg attatatggt gttatgaatg tgatgaaaaa ttatcaacgc
     1261 attgtaataa gaaggttttg gctcagatag ttgattttct ccagaaacat gcttctaaaa
     1321 cacaaacaag tgcattttct agaatcatga aactttgtga agaaaaatgt gaaacagatg
     1381 aaatacagaa gggaggaaaa tgcagaaatt tatctgtaag aggaattaca aatttaggaa
     1441 atacttgctt ttttaatgca gtcatgcaga acttggcaca gacttatact cttactgatc
     1501 tgatgaatga gatcaaagaa agtagtacaa aactcaagat ttttccttcc tcagactctc
     1561 agctggaccc attggtggtg gaactttcaa ggcctggacc actgacctca gccttgttcc
     1621 tgtttcttca cagcatgaag gagactgaaa aaggaccact ttctcctaaa gttcttttta
     1681 atcagctttg tcagaagcgg gtgcatctac atttaatatt aattaatcca actttcttgt
     1741 acaaagtggt tcgataacga attctaccgg gtaggggagg cgcttttccc aaggcagtct
     1801 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct
     1861 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc
     1921 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg
     1981 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc
     2041 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc
     2101 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc
     2161 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag
     2221 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca
     2281 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     2341 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     2401 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     2461 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     2521 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     2581 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     2641 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     2701 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     2761 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     2821 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     2881 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
     2941 catcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     3001 cctgtaggtt tggcaagcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     3061 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     3121 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     3181 cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     3241 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     3301 ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt
     3361 cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact
     3421 cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg aaattgttat
     3481 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3541 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3601 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3661 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3781 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4021 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4261 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4501 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4561 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4621 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4681 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4741 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4801 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4861 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4921 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4981 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5041 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5101 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5161 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5221 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5281 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5341 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5401 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5461 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5521 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5581 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     5641 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     5701 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     5761 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     5821 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     5881 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     5941 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     6001 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     6061 gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     6121 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     6181 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     6241 ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg
     6301 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat
     6361 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     6421 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     6481 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
     6541 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     6601 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     6661 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     6721 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     6781 agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
     6841 tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc
     6901 cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac
     6961 caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct
     7021 ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact
     7081 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
     7141 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
     7201 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     7261 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     7321 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     7381 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     7441 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     7501 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     7561 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     7621 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     7681 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     7741 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     7801 taggcgccgg aattagatcc tt
//
