LOCUS       sequence-313759-        7034 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 7034)
  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 7034)
  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-JOSD1
FEATURES             Location/Qualifiers
     source          1..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..417
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     347..369
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     385..401
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      430..439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             439..462
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             475..501
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    517..541
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1148..1172)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1185..1752
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1202..1754
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1346..1363)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1573..1592
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1766..2365
                     /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(1766..1785)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2262..2282
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2441..2955
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3117..3133
                     /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(3141..3147,3148..3165,3166..3171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3186..3207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3324..3341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3495..4083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3575..3594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4254..5045,5046..5114))
                     /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     4877..4896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5115..5219)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5287..5305
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5343..5365)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5465..5484
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5678..5700
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5828..5847
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6047..6563
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6627..6968
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
ORIGIN
        1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
       61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
      421 attagatctg ccaccatgga ctacaaggat gacgatgaca agctcgatgg aggataccca
      481 tacgatgttc cagattacgc tgctcgaggt tatcaaacaa gtttgtacaa aaaagttggc
      541 atgagttgtg tgccatggaa aggagacaag gccaaatctg aatcattgga gctgccccag
      601 gcagcacccc cacaaatcta ccatgagaaa cagcgcaggg agctttgtgc cctccacgcc
      661 ctcaataacg tcttccagga cagcaatgcc ttcacccggg atacgctgca agagattttc
      721 cagaggttgt ctccaaacac catggtgaca cctcacaaga agagcatgct gggaaatggc
      781 aactacgatg tgaatgtcat tatggcagca cttcagacca aaggctatga agctgtttgg
      841 tgggacaagc gcagggatgt cggtgtcatt gccctcacta acgtcatggg cttcatcatg
      901 aatctgccct ccagcctatg ctggggtcca ctgaaactgc ccctcaaaag gcagcactgg
      961 atctgtgttc gagaggtggg aggggcctac tacaacctcg actccaaact caagatgccc
     1021 gagtggattg gaggcgagag cgagctcagg aagtttctaa aacatcattt gcgaggaaag
     1081 aactgtgaac tcctgctggt ggtaccagaa gaggtagagg ctcatcagag ttggaggacc
     1141 gatgtgtacc caactttctt gtacaaagtg gttcgataac gaattccgcc ccccccccct
     1201 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     1261 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     1321 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     1381 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     1441 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     1501 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     1561 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     1621 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     1681 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     1741 acacgatgat aatatggcca caaccatgac cgagtacaag cccacggtgc gcctcgccac
     1801 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     1861 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     1921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     1981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     2041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     2101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     2161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     2221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     2281 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     2341 catgacccgc aagcccggtg cctgatgcat cgatgtcgac ctgcagccaa gcttatcgat
     2401 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     2461 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     2521 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     2581 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     2641 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     2701 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     2761 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     2821 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     2881 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     2941 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     3001 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     3061 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     3121 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3181 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3241 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3301 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3361 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3901 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4381 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5221 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5281 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5341 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5401 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5461 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5521 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     5581 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5641 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     5701 acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     5761 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     5821 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     5881 cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag
     5941 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     6001 catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     6061 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     6121 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     6181 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     6241 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     6301 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     6361 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     6421 gccagtcctc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct
     6481 gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact
     6541 gcccacctcg ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg
     6601 gaccaccgac ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg
     6661 tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttagcta
     6721 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
     6781 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
     6841 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
     6901 aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt
     6961 gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     7021 tgtctgaaaa ttag
//
