LOCUS       sequence-456169-        5909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hRad9 in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5909)
  AUTHORS   Lindsey-Boltz LA, Bermudez VP, Hurwitz J, Sancar A
  TITLE     Purification and characterization of human DNA damage checkpoint Rad
            complexes.
  JOURNAL   Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11236-41. doi: 
            10.1073/pnas.201373498.
  PUBMED    11572977
REFERENCE   2  (bases 1 to 5909)
  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"; doi: "10.1073/pnas"; 
            journalName: "Proc Natl Acad Sci U S A"; date: "2001-09-25- 25"; 
            volume: "98"; issue: "20"; pages: "11236-41"
COMMENT     Sequence Label: pFast-Flag-hRad9
FEATURES             Location/Qualifiers
     source          1..5909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    4..138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(41..60)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     95..114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  167..332
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      516..970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(603..622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     812..833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        996..1100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1101..1169,1170..1961)
                     /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(1319..1338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2132..2720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2621..2640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2874..2891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3025..3249
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3316..3849)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4019..4024,4025..4041,4042..4047))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        4399..4490
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4415..4432
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5718..5741
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
       61 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
      121 tcttatcatg tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga
      181 aatctagttc caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc
      241 cagttcccat ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc
      301 ctcccagttc ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt
      361 ttaatcaaac atcctgccaa ctccatgtga caaaccgtca tcttcggcta ctttttctct
      421 gtcacagaat gaaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat
      481 atcaacgctt atttgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
      541 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      601 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      661 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      721 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      781 ccctttgacg ttggagtcca cgttcttaat agtggactct tgttccaaac tggaacaaca
      841 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
      901 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
      961 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1021 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1081 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1141 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1201 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1261 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1321 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1381 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1441 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1501 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1561 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1621 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1681 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1741 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1801 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1861 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1921 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1981 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2041 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2101 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2161 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2221 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2281 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2341 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2401 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2461 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2521 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2581 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2641 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2701 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2761 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2821 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2881 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     2941 gtatttcaca ccgcatagac cagccgcgta acctggcaaa atcggttacg gttgagtaat
     3001 aaatggatgc cctgcgtaag cgggtgtggg cggacaataa agtcttaaac tgaacaaaat
     3061 agatctaaac tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag
     3121 tccagttatg ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat
     3181 tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg catggtaaag
     3241 actatattcg cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc
     3301 ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc
     3361 ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac
     3421 gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt
     3481 ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact
     3541 acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg
     3601 gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga
     3661 gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag
     3721 agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat
     3781 acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     3841 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     3901 agactgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
     3961 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
     4021 tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt
     4081 gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa
     4141 cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta
     4201 cggcaaggtg ctgtgcacgg atctgccctg gcttcaggag atcggaagac ctcggccgtc
     4261 gcggcgcttg ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga
     4321 aggcgagcat cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat
     4381 actccggaat attaatagat catggagata attaaaatga taaccatctc gcaaataaat
     4441 aagtatttta ctgttttcgt aacagttttg taataaaaaa acctataaat attccggatt
     4501 attcataccg tcccaccatc gggcgcggat ccaccatgaa gtgcctggtc acgggcggca
     4561 acgtgaaggt gctcggcaag gccgtccact ccctgtcccg catcggggac gagctctacc
     4621 tggaaccctt ggaggacggg ctctccctcc ggacggtgaa ctcctcccgc tctgcctatg
     4681 cctgctttct ctttgccccg ctcttcttcc agcaatacca ggcagccacc cctggtcagg
     4741 acctgctgcg ctgtaagatc ctgatgaagt ctttcctgtc tgtcttccgc tcactggcga
     4801 tgctggagaa gacggtggaa aaatgctgca tctccctgaa tggccggagc agccgcctgg
     4861 tggtccagct gcattgcaag ttcggggtgc ggaagactca caacctgtcc ttccaggact
     4921 gtgagtccct gcaggccgtc ttcgacccag cctcgtgccc ccacatgctc cgcgccccag
     4981 cacgggttct gggggaggct gttctgccct tctctcctgc actggctgaa gtgacgctgg
     5041 gcattggccg tggccgcagg gtcatcctgc gcagctacca cgaggaggag gcagacagca
     5101 ctgccaaagc catggtgact gagatgtgcc ttggagagga ggatttccag cagctgcagg
     5161 cccaggaagg ggtggccatc actttctgcc tcaaggaatt ccgggggctc ctgagctttg
     5221 cagagtcagc aaacttgaat cttagcattc attttgatgc tccaggcagg cccgccatct
     5281 tcaccatcaa ggactctttg ctggacggcc actttgtctt ggccacactc tcagacaccg
     5341 actcgcactc ccaggacctg ggctccccag agcgtcacca gccagtgcct cagctccagg
     5401 ctcacagcac accccacccg gacgactttg ccaatgacga cattgactct tacatgatcg
     5461 ccatggaaac cactataggc aatgagggct cgcgggtgct gccctccatt tccctttcac
     5521 ctggccccca gccccccaag agccccggtc cccactccga ggaggaagat gaggctgagc
     5581 ccagtacagt gcctgggact cccccaccca agaagttccg ctcactgttc ttcggctcca
     5641 tcctggcccc tgtacgctcc ccccagggcc ccagccctgt gctggcggaa gacagtgagg
     5701 gtgaagtcga caccatggat tacaaagacg atgacgacaa ggctagctag tctagagcct
     5761 gcagtctcga ggcatgcggt accaagcttg tcgagaagta ctagaggatc ataatcagcc
     5821 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     5881 tgaaacataa aatgaatgca attgttgtt
//
