LOCUS       sequence-275985-        9140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  codon optimized expression and purification of human FANCD2 in 
            insect cells using baculovirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9140)
  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 9140)
  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: pFastbac1-Flag-FANCD2
FEATURES             Location/Qualifiers
     source          1..9140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..455
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     297..318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        481..585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(586..654,655..1446)
                     /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(804..823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1617..2205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2106..2125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2359..2376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2510..2734
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2801..3334)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3523..3528,3529..3545,3546..3551))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3903..3994
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3919..3936
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4096..4119
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    8629..8763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8666..8685)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8720..8739
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8792..8957
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc ttaatagtgg
      301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
      361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
      421 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
      481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1741 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2281 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2341 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2401 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagaccagcc gcgtaacctg
     2461 gcaaaatcgg ttacggttga gtaataaatg gatgccctgc gtaagcgggt gtgggcggac
     2521 aataaagtct taaactgaac aaaatagatc taaactatga caataaagtc ttaaactaga
     2581 cagaatagtt gtaaactgaa atcagtccag ttatgctgtg aaaaagcata ctggactttt
     2641 gttatggcta aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag
     2701 gggcgtggcc aagggcatgg taaagactat attcgcggcg ttgtgacaat ttaccgaaca
     2761 actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg gtacttgggt
     2821 cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga gccactgcgg
     2881 gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg ttggcctcat
     2941 gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg ccggagactg
     3001 cgagatcata gatatagatc tcactacgcg gctgctcaaa cttgggcaga acgtaagccg
     3061 cgagagcgcc aacaaccgct tcttggtcga aggcagcaag cgcgatgaat gtcttactac
     3121 ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg gctacgtctc
     3181 cgaactcacg accgaaaaga tcaagagcag cccgcatgga tttgacttgg tcagggccga
     3241 gcctacatgt gcgaatgatg cccatacttg agccacctaa ctttgtttta gggcgactgc
     3301 cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc tgctccataa catcaaacat
     3361 cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg tacaaaaaaa
     3421 cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag
     3481 gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagtttacg aaccgaacag
     3541 gcttatgtca actgggttcg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc
     3601 ttgggcagca gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg caaggtttcg
     3661 gtctccacgc atcgtcaggc attggcggcc ttgctgttct tctacggcaa ggtgctgtgc
     3721 acggatctgc cctggcttca ggagatcgga agacctcggc cgtcgcggcg cttgccggtg
     3781 gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg
     3841 ttcgcccagg actctagcta tagttctagt ggttggctac gtatactccg gaatattaat
     3901 agatcatgga gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt
     3961 tcgtaacagt tttgtaataa aaaaacctat aaatattccg gattattcat accgtcccac
     4021 catcgggcgc ggatcccggt ccgaagcgcg cggaattcaa aggcctacgt cgacgagctc
     4081 ctagctagct agatggacta caaagacgat gacgacaagg gagcagtgtc taaacgccgc
     4141 ctctcaaaat cagaagataa agaaagcctc accgaagatg cctcaaaaac ccgtaaacag
     4201 ccactctcaa agaagaccaa gaagagccac atcgccaacg aggtggagga aaacgactct
     4261 atcttcgtga agctgctgaa gatcagcgga atcatcctga agaccggtga atcccagaac
     4321 cagctggccg tggaccagat cgctttccag aagaagctgt tccagaccct gcgcaggcac
     4381 cctagctacc caaagatcat cgaggagttc gtgagcggcc tggagtctta catcgaggac
     4441 gaagactctt tcagaaactg cctgctgagc tgcgaacgcc tgcaggacga ggaagcctcc
     4501 atgggagcca gctactccaa gtctctgatc aagctgctgc tgggtatcga catcctgcag
     4561 cccgctatca tcaagaccct gttcgaaaag ctgcctgagt acttcttcga aaacaagaac
     4621 tctgacgaga tcaacatccc ccgcctgatc gtgagccagc tgaagtggct ggacagggtg
     4681 gtggacggca aggacctgac caccaagatc atgcagctga tctccatcgc cccagagaac
     4741 ctgcagcacg acatcatcac cagcctgccc gaaatcctgg gagactctca gcacgctgac
     4801 gtgggcaagg agctgagcga cctgctgatc gaaaacacca gcctgaccgt gcctatcctg
     4861 gacgtgctga gcagcctgcg cctggaccca aacttcctgc tgaaggtgag gcagctggtc
     4921 atggacaagc tgtctagcat ccgcctggag gacctgcctg tgatcatcaa gttcatcctg
     4981 cactccgtga ccgccatgga caccctggaa gtgatctctg agctgagaga aaagctggac
     5041 ctgcagcact gcgtgctgcc atcccgcctg caggcctctc aggtgaagct gaagagcaag
     5101 ggcagggctt cctctagcgg aaaccaggag tcctctggtc agtcctgcat catcctgctg
     5161 ttcgacgtga tcaagtctgc catccgctac gagaagacca tcagcgaagc ctggatcaag
     5221 gctatcgaga acaccgccag cgtgagcgaa cacaaggtgt tcgacctggt catgctgttc
     5281 atcatctact ccaccaacac ccagaccaag aagtacatcg acagggtgct gagaaacaag
     5341 atccgctctg gctgcatcca ggagcagctg ctgcagtcca ccttctctgt gcactacctg
     5401 gtgctgaagg acatgtgcag ctccatcctg agcctggccc agtccctgct gcactctctg
     5461 gaccagagca tcatctcctt cggatctctg ctgtacaagt acgccttcaa gttcttcgac
     5521 acctactgcc agcaggaagt ggtgggtgct ctggtgaccc acatctgcag cggcaacgag
     5581 gccgaagtgg acaccgctct ggacgtgctg ctggaactgg tggtgctgaa cccatctgcc
     5641 atgatgatga acgccgtgtt cgtgaagggt atcctggact acctggacaa catcagcccc
     5701 cagcagatcc gcaagctgtt ctacgtgctg agcaccctgg ccttctccaa gcagaacgag
     5761 gcttctagcc acatccagga cgacatgcac ctggtcatcc gcaagcagct gtcctctacc
     5821 gtgttcaagt acaagctgat cggcatcatc ggagccgtga ccatggctgg catcatggcc
     5881 gctgacagat ctgagagccc ctccctgacc caggaacgcg ccaacctgtc cgacgagcag
     5941 tgcacccagg tgacctctct gctgcagctg gtgcactctt gttccgagca gagcccccag
     6001 gcctccgctc tgtactacga cgagttcgcc aacctgatcc agcacgagaa gctggaccct
     6061 aaggctctgg aatgggtggg ccacaccatc tgcaacgact tccaggacgc cttcgtggtg
     6121 gacagctgcg tggtgcctga gggagacttc cccttccctg tgaaggctct gtacggactg
     6181 gaggaatacg acacccagga cggtatcgcc atcaacctgc tgcccctgct gttcagccag
     6241 gacttcgcta aggacggcgg acctgtgacc agccaggagt ccggacagaa gctggtgagc
     6301 cccctgtgcc tggcccctta cttccgcctg ctgaggctgt gcgtggaacg ccagcacaac
     6361 ggtaacctgg aggaaatcga cggcctgctg gactgcccta tcttcctgac cgacctggag
     6421 ccaggtgaaa agctggagtc catgtctgcc aaggagagat ctttcatgtg cagcctgatc
     6481 ttcctgaccc tgaactggtt ccgcgaaatc gtgaacgctt tctgccagga gaccagccct
     6541 gaaatgaagg gaaaggtgct gacccgcctg aagcacatcg tggagctgca gatcatcctg
     6601 gaaaagtacc tggctgtgac cccagactac gtgccccctc tgggtaactt cgacgtggag
     6661 accctggaca tcacccctca caccgtgacc gccatctctg ctaagatccg caagaagggc
     6721 aagatcgaaa ggaagcagaa gaccgacggc agcaagacca gctcctctga caccctgtct
     6781 gaggaaaaga acagcgagtg tgacccaacc cctagccaca ggggacagct gaacaaggag
     6841 ttcaccggca aggaagagaa gaccagcctg ctgctgcaca actcccacgc cttcttcagg
     6901 gagctggaca tcgaagtgtt ctccatcctg cactgcggtc tggtgaccaa gttcatcctg
     6961 gacaccgaga tgcacaccga agctaccgag gtggtgcagc tgggcccacc cgagctgctg
     7021 ttcctgctgg aagacctgtc ccagaagctg gagtctatgc tgacccctcc aatcgccaga
     7081 cgcgtgccat tcctgaagaa caagggttct aggaacatcg gcttcagcca cctgcagcag
     7141 agatccgctc aggagatcgt gcactgcgtg ttccagctgc tgacccctat gtgcaaccac
     7201 ctggagaaca tccacaacta cttccagtgc ctggccgctg aaaaccacgg cgtggtggac
     7261 ggtcctggcg tgaaggtgca ggagtaccac atcatgagca gctgctacca gcgcctgctg
     7321 cagatcttcc acggcctgtt cgcctggtct ggattcagcc agccagagaa ccagaacctg
     7381 ctgtactccg ctctgcacgt gctgtctagc cgcctgaagc agggagaaca ctctcagcct
     7441 ctggaggaac tgctgagcca gtccgtgcac tacctgcaga acttccacca gtccatccca
     7501 tctttccagt gcgccctgta cctgatccgc ctgctgatgg tcatcctgga gaagagcacc
     7561 gcctccgctc agaacaagga aaagatcgcc tctctggcta ggcagttcct gtgcagagtg
     7621 tggcccagcg gagacaagga gaagtctaac atcagcaacg accagctgca cgctctgctg
     7681 tgcatctacc tggaacacac cgagagcatc ctgaaggcca tcgaggaaat cgctggagtg
     7741 ggtgtgccag agctgatcaa ctcccccaag gacgcctcct ctagcacctt cccaaccctg
     7801 accaggcaca ccttcgtggt gttcttcaga gtgatgatgg ctgagctgga aaagaccgtg
     7861 aagaagatcg agcctggaac tgctgctgac tcccagcaga tccacgagga aaagctgctg
     7921 tactggaaca tggccgtgcg cgacttcagc atcctgatca acctgatcaa ggtgttcgac
     7981 tcccaccctg tgctgcacgt gtgcctgaag tacggtcgcc tgttcgtgga ggctttcctg
     8041 aagcagtgca tgccactgct ggacttcagc ttcagaaagc accgcgaaga cgtgctgtcc
     8101 ctgctggaga ccttccagct ggacaccagg ctgctgcacc acctgtgcgg ccacagcaag
     8161 atccaccagg acaccagact gacccagcac gtgcctctgc tgaagaagac cctggagctg
     8221 ctggtgtgca gagtgaaggc catgctgacc ctgaacaact gccgcgaagc tttctggctg
     8281 ggcaacctga agaacaggga cctgcaggga gaggaaatca agtcccagaa ctctcaggag
     8341 agcaccgccg acgagtccga agacgacatg tcctctcagg cctctaagag caaggctacc
     8401 gaagacggcg aggaagacga agtctctgct ggtgaaaaag aacaggacag tgacgaaagt
     8461 tatgatgaca gtgattgacc gctcgaggca tgcggtacca agcttgtcga gaagtactag
     8521 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     8581 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     8641 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     8701 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     8761 atctgatcac tgcttgagcc taggagatcc gaaccagata agtgaaatct agttccaaac
     8821 tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt
     8881 ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac
     8941 tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat caaacatcct
     9001 gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac agaatgaaaa
     9061 tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa cgcttatttg
     9121 cagcctgaat ggcgaatggg
//
