LOCUS       sequence-288208-        7639 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 7639)
  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 7639)
  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..7639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /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     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /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
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     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    4160..4208
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4233..4252
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(6625..6648,6649..6684,6685..6708))
                     /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(6746..6855)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6879..6970
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6895..6912
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    7229..7363
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7266..7285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7320..7339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7392..7557
                     /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 cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc 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 acttgggcag 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 cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
     4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
     4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
     4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4261 cccccatctc ccggtaccgc atgcttagga gtcgaggggc agcagttcga aggacatgac
     4321 gcagtcgatg aaccaggagg aaggagcctt ccacactttg ccctgacgca cacgctcagg
     4381 atggtagttg cacaggtcct cgtagatgat gtactgggtg cagaatcttt ggtcggagtc
     4441 gggacgagcg tggtaagcca cagtgttgat ggtctgggtc acgtcggagt caggcttagg
     4501 cttcctagac aggatctggc caccaccagc ggtcacgagc ttgatcaggt tgtccttagg
     4561 atggtgcttg aaggtgcccc acaggtagaa gtagcagccg tcgaacagct tggggagcag
     4621 ctgttcacgg ttcagacgag aacgacgggg accctcgggg atctcgtact tctcttcttg
     4681 ttcgcacact ttacgacgca ggcaagcctt gacccactcg aacttgagta tccagcaacc
     4741 gttcaggata cccagcatac acttcagggt agactgcaca gcgtcgcctg gaaccaccac
     4801 gtgagtcacg gtgctgtcga attcggtgta cttcttagcc ttcaggatca cagccagctc
     4861 ggacagcatc ttttgctgct cggaggacag tccagaaccg atcaacacca ggggaccgtc
     4921 acgacgttga ccggtgttca tcacggagca gtgggaagca gaggaggact cgttcttctc
     4981 gggcaacagc agcagggact tcatggactc gtcgtcggtg tagtccacag gacgcaggcc
     5041 gaagatgttc acagcgttac gggaagcacc gtaggacagc aacagcttca cgatgtccac
     5101 gtgaccgttc ttagcagcgt cgtgcagggg agagtcgttc tggtatccgg tggtgttgac
     5161 cagagccttg tgttgcagca gcagctcgac caccttcagg tgaccatggt tacaagcctc
     5221 gtgcagagga gtccatccag cgtggtcctt cacgttgggg tcagaaccgt tttggagcag
     5281 gtactcgacg gaggggatgt cgcccttgat agaagcaatg tgcagcagag tctcgccacg
     5341 gtggttacgc ttcacagcca tgttgggcag cagcttcata gcggatggag aggacatcac
     5401 acgacggtaa gaggaggagg acagagttga aggaggagta ccgggagaca gagagatgaa
     5461 ctcgtcggac atgttggagt tcttgcgacc ggaagtacca ccgactttac gcttgcagga
     5521 aggtggagag gagcactcag gcagagggat gttctcggaa ggcacggtct gcttcacgaa
     5581 gtcgccagag gtggacagga tggacgtccg gcaacgcttg gagataggtg aggacagacg
     5641 gttgtggtga ccacgcttgc cgttgttttc caggggcaga gacttcttgg aggtcaggta
     5701 gttcttgcac actttctcag gggtgaccac ctcgttacga gacttggtgt caggggactc
     5761 gatctgctca gccagaggca gggacacttc ggtcagggaa ccgaagcact cagactcggt
     5821 cagagaaccg gaagccagca ggtcgatttc gccgttgatc tgaggggagc tgatcacgga
     5881 aggctggctg cagaagctga cgagcttttg cttggattcc tctttggagt cgaactcgcc
     5941 gtcctctttt tcagcttcga ggttccactt ctggttgatc tcagccaggg ttttcttctt
     6001 ctgcttcttg ccggaacgag cagaagcctt cttagcacgc tcggacacgt cagcaggagg
     6061 agaaggggac acgaactcgt aggagtcctg ctgagcgcta gcgtccttct tgatagcggg
     6121 ctgagtctgc acggaagcct tggacaccac gtagcgcact ttcttggagc ggggcgagaa
     6181 ccacatcttg atggagtttt tcttgttgcc agcgtcgttg aacagggact tgcggggctt
     6241 gtcctctttc aggtcggaca gctcgttgtc gtgcagcagg ttcctcagct tggagcacag
     6301 ttggatcatg gagtccagct ggcggttgat cttcaggtcc tgtatccaag caggggtgta
     6361 gcacacgggg caaccagtac cgatgcagtc ggacacgcag ttggagcaga agatgtgctc
     6421 gcaaccaccg aggcacacag gttcacgcag gatgttggtg caacgggagc acctcagcag
     6481 cttttccaga cggtccagag cagcacgaga gtgagcccaa gcaccacgac catcaggttc
     6541 catagcagga gcggaacgag gctcgtttcc ggaacggata cgtggctgcc tgttacgagg
     6601 ctgacggttg tcaggcatcc cgggcttctc aaattgtggg tgtgaccatg ctgaaccacc
     6661 tgagcctcct ccagagccac ctcccttctc gaactgtggg tggctccaca tctcgagatc
     6721 ccgggtgatc aagtcttcgt cgagtgattg taaataaaat gtaatttaca gtatagtatt
     6781 ttaattaata tacaaatgat ttgataataa ttcttattta actataatat attgtgttgg
     6841 gttgaattaa aggtccgtat actccggaat attaatagat catggagata attaaaatga
     6901 taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa
     6961 acctataaat attccggatt attcataccg tcccaccatc gggcgcggat cccggtccga
     7021 agcgcgcgga attcaaaggc ctacgtcgac gagctcacta gtcgcggccg ctttcgaatc
     7081 tagagcctgc agtctcgaca agcttgtcga gaagtactag aggatcataa tcagccatac
     7141 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     7201 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     7261 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7321 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac tgcttgagcc
     7381 taggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt
     7441 tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata
     7501 atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc gcccacagcg
     7561 gggcattttt cttcctgtta tgtttttaat caaacatcct gccaactcca tgtgacaaac
     7621 cgtcatcttc ggctacttt
//
