LOCUS       sequence-270608-        7640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human StrepII-tagged BARD1 for purification from 
            baculovirus/insect cell system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7640)
  AUTHORS   Tan W, Murphy VJ, Charron A, van Twest S, Sharp M, Constantinou A, 
            Parker MW, Crismani W, Bythell-Douglas R, Deans AJ
  TITLE     Preparation and purification of mono-ubiquitinated proteins using 
            Avi-tagged ubiquitin.
  JOURNAL   PLoS One. 2020 Feb 24;15(2):e0229000. doi: 
            10.1371/journal.pone.0229000. eCollection 2020.
  PUBMED    32092106
REFERENCE   2  (bases 1 to 7640)
  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: "PLoS One.";
            date: "2020-02-24"; volume: "15(2):e0229000. doi"; pages: " 
            10.1371/journal.pone.0229000. eCollection 2020"
COMMENT     Sequence Label: pFastbac-Dual-StrepII-BARD1
FEATURES             Location/Qualifiers
     source          1..7640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     399..420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        584..688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(689..757,758..1549)
                     /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(907..926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1720..2308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2209..2228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2462..2479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4161..4209
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4234..4253
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(6626..6649,6650..6685,6686..6709))
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     promoter        complement(6747..6856)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6880..6971
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6896..6913
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    7230..7364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7267..7286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7321..7340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7393..7558
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      541 attaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
      961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2101 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2461 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2521 tctgtgcggt atttcacacc gcagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcct acgtacccgt agtggctatg gcagggcttg ccgccccgac
     4021 gttggctgcg agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga
     4081 aacgcgggcg tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg
     4141 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt
     4201 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     4261 tcccccatct cccggtaccg catgcttagg agtcgagggg cagcagttcg aaggacatga
     4321 cgcagtcgat gaaccaggag gaaggagcct tccacacttt gccctgacgc acacgctcag
     4381 gatggtagtt gcacaggtcc tcgtagatga tgtactgggt gcagaatctt tggtcggagt
     4441 cgggacgagc gtggtaagcc acagtgttga tggtctgggt cacgtcggag tcaggcttag
     4501 gcttcctaga caggatctgg ccaccaccag cggtcacgag cttgatcagg ttgtccttag
     4561 gatggtgctt gaaggtgccc cacaggtaga agtagcagcc gtcgaacagc ttggggagca
     4621 gctgttcacg gttcagacga gaacgacggg gaccctcggg gatctcgtac ttctcttctt
     4681 gttcgcacac tttacgacgc aggcaagcct tgacccactc gaacttgagt atccagcaac
     4741 cgttcaggat acccagcata cacttcaggg tagactgcac agcgtcgcct ggaaccacca
     4801 cgtgagtcac ggtgctgtcg aattcggtgt acttcttagc cttcaggatc acagccagct
     4861 cggacagcat cttttgctgc tcggaggaca gtccagaacc gatcaacacc aggggaccgt
     4921 cacgacgttg accggtgttc atcacggagc agtgggaagc agaggaggac tcgttcttct
     4981 cgggcaacag cagcagggac ttcatggact cgtcgtcggt gtagtccaca ggacgcaggc
     5041 cgaagatgtt cacagcgtta cgggaagcac cgtaggacag caacagcttc acgatgtcca
     5101 cgtgaccgtt cttagcagcg tcgtgcaggg gagagtcgtt ctggtatccg gtggtgttga
     5161 ccagagcctt gtgttgcagc agcagctcga ccaccttcag gtgaccatgg ttacaagcct
     5221 cgtgcagagg agtccatcca gcgtggtcct tcacgttggg gtcagaaccg ttttggagca
     5281 ggtactcgac ggaggggatg tcgcccttga tagaagcaat gtgcagcaga gtctcgccac
     5341 ggtggttacg cttcacagcc atgttgggca gcagcttcat agcggatgga gaggacatca
     5401 cacgacggta agaggaggag gacagagttg aaggaggagt accgggagac agagagatga
     5461 actcgtcgga catgttggag ttcttgcgac cggaagtacc accgacttta cgcttgcagg
     5521 aaggtggaga ggagcactca ggcagaggga tgttctcgga aggcacggtc tgcttcacga
     5581 agtcgccaga ggtggacagg atggacgtcc ggcaacgctt ggagataggt gaggacagac
     5641 ggttgtggtg accacgcttg ccgttgtttt ccaggggcag agacttcttg gaggtcaggt
     5701 agttcttgca cactttctca ggggtgacca cctcgttacg agacttggtg tcaggggact
     5761 cgatctgctc agccagaggc agggacactt cggtcaggga accgaagcac tcagactcgg
     5821 tcagagaacc ggaagccagc aggtcgattt cgccgttgat ctgaggggag ctgatcacgg
     5881 aaggctggct gcagaagctg acgagctttt gcttggattc ctctttggag tcgaactcgc
     5941 cgtcctcttt ttcagcttcg aggttccact tctggttgat ctcagccagg gttttcttct
     6001 tctgcttctt gccggaacga gcagaagcct tcttagcacg ctcggacacg tcagcaggag
     6061 gagaagggga cacgaactcg taggagtcct gctgagcgct agcgtccttc ttgatagcgg
     6121 gctgagtctg cacggaagcc ttggacacca cgtagcgcac tttcttggag cggggcgaga
     6181 accacatctt gatggagttt ttcttgttgc cagcgtcgtt gaacagggac ttgcggggct
     6241 tgtcctcttt caggtcggac agctcgttgt cgtgcagcag gttcctcagc ttggagcaca
     6301 gttggatcat ggagtccagc tggcggttga tcttcaggtc ctgtatccaa gcaggggtgt
     6361 agcacacggg gcaaccagta ccgatgcagt cggacacgca gttggagcag aagatgtgct
     6421 cgcaaccacc gaggcacaca ggttcacgca ggatgttggt gcaacgggag cacctcagca
     6481 gcttttccag acggtccaga gcagcacgag agtgagccca agcaccacga ccatcaggtt
     6541 ccatagcagg agcggaacga ggctcgtttc cggaacggat acgtggctgc ctgttacgag
     6601 gctgacggtt gtcaggcatc ccgggcttct caaattgtgg gtgtgaccat gctgaaccac
     6661 ctgagcctcc tccagagcca cctcccttct cgaactgtgg gtggctccac atctcgagat
     6721 cccgggtgat caagtcttcg tcgagtgatt gtaaataaaa tgtaatttac agtatagtat
     6781 tttaattaat atacaaatga tttgataata attcttattt aactataata tattgtgttg
     6841 ggttgaatta aaggtccgta tactccggaa tattaataga tcatggagat aattaaaatg
     6901 ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt gtaataaaaa
     6961 aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga tcccggtccg
     7021 aagcgcgcgg aattcaaagg cctacgtcga cgagctcact agtcgcggcc gctttcgaat
     7081 ctagagcctg cagtctcgac aagcttgtcg agaagtacta gaggatcata atcagccata
     7141 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     7201 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     7261 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     7321 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
     7381 ctaggagatc cgaaccagat aagtgaaatc tagttccaaa ctattttgtc atttttaatt
     7441 ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc ttccctaaat
     7501 aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc cgcccacagc
     7561 ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc tgccaactcc atgtgacaaa
     7621 ccgtcatctt cggctacttt
//
