LOCUS       sequence-129563-        7700 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 7700)
  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 7700)
  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-STAMBP
FEATURES             Location/Qualifiers
     source          1..7700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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)."
     LTR             307..821
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(897..1496)
                     /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(980..1000)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1477..1496
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    1508..2060
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1510..2077
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1670..1689)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1899..1916
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    2090..2114
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3387..3411)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3427..3453)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3466..3489)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3489..3498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3511..3927)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3527..3543)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3559..3581)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3994..4335
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4399..4915
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5115..5134)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5262..5284)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5478..5497)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5597..5619
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5657..5675)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5743..5847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5848..5916,5917..6708)
                     /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     complement(6066..6085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6879..7467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7368..7387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7621..7638
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc
      181 ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc
      241 aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg
      301 ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      361 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      421 gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt
      481 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      541 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      601 gaaactgctg agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc
      661 aggaactgct taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa
      721 ccttgatctg aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt
      781 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
      841 ctaaataaaa tcttttattt tatcgataag cttggctgca ggtcgacatc gatgcatcag
      901 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
      961 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
     1021 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
     1081 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
     1141 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
     1201 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     1261 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     1321 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     1381 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     1441 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtt
     1501 gtggccatat tatcatcgtg tttttcaaag gaaaaccacg tccccgtggt tcggggggcc
     1561 tagacgtttt tttaacctcg actaaacaca tgtaaagcat gtgcaccgag gccccagatc
     1621 agatcccata caatggggta ccttctgggc atccttcagc cccttgttga atacgcttga
     1681 ggagagccat ttgactcttt ccacaactat ccaactcaca acgtggcact ggggttgtgc
     1741 cgcctttgca ggtgtatctt atacacgtgg cttttggccg cagaggcacc tgtcgccagg
     1801 tggggggttc cgctgcctgc aaagggtcgc tacagacgtt gtttgtcttc aagaagcttc
     1861 cagaggaact gcttccttca cgacattcaa cagaccttgc attcctttgg cgagagggga
     1921 aagaccccta ggaatgctcg tcaagaagac agggccaggt ttccgggccc tcacattgcc
     1981 aaaagacggc aatatggtgg aaaataacat atagacaaac gcacaccggc cttattccaa
     2041 gcggcttcgg ccagtaacgt tagggggggg gggcggaatt cgttatcgaa ccactttgta
     2101 caagaaagtt gggcatcgaa ggtctgtgat ggtcactgct ctgtccacaa cagtcacgtg
     2161 gctgcagcta cagaacagag gtggatcctt gctgtgtgga tgaaatcctt tctggcgaca
     2221 ggaagaaatc tcctctagtc catggtcagt tagtttaaag aatccagttt cctggaactt
     2281 gggggagcaa acaatggcta ctgactctgg caacatcatc tggtaagagc agtgagtgtg
     2341 taggtcgaca ctggagagaa acgcggtctg tgtggggtga gtatgaatcc agcccagtgt
     2401 gatgaggccc tgctgatcct gtatgaggaa aagttcttct tcgttctctg tgttgcagta
     2461 atcagaccca gcactttgct tggggatgag aacatgggta atggtaaatt cattcctcat
     2521 cagttttcca cagagaattc cacatgtctc cactccccgg gcagtgttgg cactggctaa
     2581 ctggagaaac tgtgggcaca gccgcccagg caccaccaca tggcgcaatc catcgattgt
     2641 gggaatactt tctgagttgc tcagtgctcc aggtttcaag gacctgtcca ccacaggtgg
     2701 cttagctggc cttacagttg tgtgacagtc tgaaggctgt atggatgaga ctgttaaggt
     2761 ggggaacaca tctaaggagg gcttctccaa gtcaggcact agcgggccac ctaggccagg
     2821 gtctaccttc ccaaactcct gtacaatttt cagtcgctct ttttctagct cctggttccg
     2881 gatcatctcc tcgaaggcat ggaactgttc ctgttccaat tgctgctgct tctgttgtgc
     2941 taccctctgt ttttcctttt ccagctcttg ctggatggcc atgttccggg ccaattcctc
     3001 tgcttccttc ttcttttctt cattatattc tgtatattct ttggtatatc gttttaacag
     3061 ctctgccttc agctcttctg ctttgggaaa tgcaatctcc tttaatttct ttactgtgtc
     3121 tttcttttca ggaatgacag cagatttgta atctcgatgt tttggtagtt tctcaataaa
     3181 gagcgtgata tacttgttat agaggatgaa ggcatgttca atgttgcctt cctcagagta
     3241 aatggatgcc attcggataa tctcaactcc agagcggaag taccgacggg gtggaatgtc
     3301 ttcattcacc tctaccgcac tacccagctg ggagagagcc ctcacccggt cttcgggcgg
     3361 gaggctcaca tctccgtggt cagacatgcc aacttttttg tacaaacttg tttgataacc
     3421 tcgagcagcg taatctggaa catcgtatgg gtatcctcca tcgagcttgt catcgtcatc
     3481 cttgtagtcc atggtggcag atctaattcc ggcgcctaga gaaggagtga gggctggata
     3541 aagggaggat cgaggcgggg tcgaacgagg aggttcaagg gggagagacg gggcggatgg
     3601 aggaagagga ggcggaggct tagggtgtac aaagggcttg acccagggag gggggtcaaa
     3661 agccaaggct tcccaggtca cgatgtaggg gacctggtct gggtgtccat gcgggccagg
     3721 tgaaaagacc ttgatcttaa cctgggtgat gaggtctcgg ttaaaggtgc cgtctcgcgg
     3781 ccatccgacg ttaaaggttg gccattctgc agagcagaag gtaacccaac gtctcttctt
     3841 gacatctacc gactggttgt gagcgatccg ctcgacatct ttccagtgac ctaaggtcaa
     3901 acttaaggga gtggtaacag tctggcccta attttcagac aaatacagaa acacagtcag
     3961 acagagacaa cacagaacga tgctgcagcg ctgcagcaga caagacgcgc ggcttcggtt
     4021 ccaaaccgaa agcaaaaatt cagacggagg cgggaactgt tttaggttct cgtctcctac
     4081 cagaaccaca tatcctgacg gggtcggatt ccacatcgac tcccttcctc aggtcgggcc
     4141 acaaaaacgg cccccaaagt ccctgggacg tctcccaggg ttgcggccgg gtgttcagaa
     4201 ctcgtcagtt ccaccacggg tccgccagat acagagctag ttagctaact agtacagacg
     4261 caggcgcaaa cattagatgc cggcacaaac acgcacaaag acagagacag acacgaaacg
     4321 accgctggcc agcttacctc ccggcggggg ggtcggtggt ccctgggcag gggtctccaa
     4381 atctcggtgg aacctccaaa tgaaagaccc ccgaggtggg cagtcaatca atctgaggag
     4441 accctcccaa ggatcagcga gtccacgatt cggatgcaaa cagcaagagg ctttattggg
     4501 aatacgggta cccgggcgac gcagtctatc ggaggactgg cgcgccgagt gaggggttgt
     4561 gggctctttt attgagctcg gggagcagaa gcgcgcgaac agaagcgaga agcgaactga
     4621 ttggttagtt caaataaggc acagggtcat ttcaggtcct tggggcaccc tggaaacatc
     4681 tgatggttct ctagaaactg ctgagggcgg gaccgcatct ggggaccatc tgttcttggc
     4741 cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata ttctgctgtc
     4801 tctctgttcc taaccttgat ctgaacttct ctattctcag ttatgtattt tccatgcctt
     4861 gcaaaatggc gttacttaag ctagcttgcc aaacctacag gtggggtctt tcattccccc
     4921 ctttttctgg agactaaata aaatctttta ttttatctat ggctcgtact ctataggctt
     4981 cagctggtga tattgttgag tcaaaactag agcctggacc actgatatcc tgtctttaac
     5041 aaattggact aatcgcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     5101 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     5161 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     5221 gttgggcgcc atctccttgc atgcaccatt ccttgcgccg tcgttttaca acgtcgtgac
     5281 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5341 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5401 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5461 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5521 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5581 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5641 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5701 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5761 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5821 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5881 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5941 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6001 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6061 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6121 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6181 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6241 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6301 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6361 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6421 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6481 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6541 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6601 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6661 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6721 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6781 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6841 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6901 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6961 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7021 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7081 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7141 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7201 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7261 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7321 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7381 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7441 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7501 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7561 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7621 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     7681 cgttggccga ttcattaatg
//
