LOCUS       sequence-456180-        6851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hRad17 in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6851)
  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 6851)
  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-His6-hRad17
FEATURES             Location/Qualifiers
     source          1..6851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    475..609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(512..531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     566..585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  638..803
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      987..1441
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1074..1093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1283..1304
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1467..1571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1572..1640,1641..2432)
                     /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(1790..1809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2603..3191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3092..3111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3345..3362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3496..3720
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3787..4320)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4490..4495,4496..4512,4513..4518))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        4870..4961
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4886..4903
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5028..5045
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5067..5087
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ttgaaaatgg aagccctgac tgacagggaa catggaatga tagaccctga cagcggagat
       61 gaagcccagc ttaatggagg acattctgca gaggaatctc tgggtgaacc cactcaagcc
      121 actgtgccgg aaacctggtc tcttcctttg agtcagaata gtgccagtga actgcctgct
      181 agccagcccc agcccttttc agcccaagga gacatggaag aaaacataat aatagaagac
      241 tacgagagtg atgggacata gggaattcaa aggcctacgt cgacgagctc aactagtgcg
      301 gccgctttcg aatctagagc ctgcagtctc gaggcatgcg gtaccaagct tgtcgagaag
      361 tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
      421 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
      481 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      541 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
      601 gtctggatct gatcactgct tgagcctagg agatccgaac cagataagtg aaatctagtt
      661 ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca
      721 tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt
      781 cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa
      841 catcctgcca actccatgtg acaaaccgtc atcttcggct actttttctc tgtcacagaa
      901 tgaaaatttt tctgtcatct cttcgttatt aatgtttgta attgactgaa tatcaacgct
      961 tatttgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     1021 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1081 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1141 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1201 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1261 gttggagtcc acgttcttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1321 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1381 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1441 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1501 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1561 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1621 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1681 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1741 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1801 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1861 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1921 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1981 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2041 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2101 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2161 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2221 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2281 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2341 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2401 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2461 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2521 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2581 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2641 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2701 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2761 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2821 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2881 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2941 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3001 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3061 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3241 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3301 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3361 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3421 accgcataga ccagccgcgt aacctggcaa aatcggttac ggttgagtaa taaatggatg
     3481 ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa
     3541 ctatgacaat aaagtcttaa actagacaga atagttgtaa actgaaatca gtccagttat
     3601 gctgtgaaaa agcatactgg acttttgtta tggctaaagc aaactcttca ttttctgaag
     3661 tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg gcatggtaaa gactatattc
     3721 gcggcgttgt gacaatttac cgaacaactc cgcggccggg aagccgatct cggcttgaac
     3781 gaattgttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc
     3841 caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac
     3901 ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc
     3961 ctgcctccgg tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg
     4021 ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc
     4081 agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg
     4141 atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg
     4201 catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc
     4261 acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     4321 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     4381 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc
     4441 ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac
     4501 cgaacaggct tatgtcaact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc
     4561 ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa
     4621 ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt
     4681 gctgtgcacg gatctgccct ggcttcagga gatcggaaga cctcggccgt cgcggcgctt
     4741 gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca
     4801 tcgtttgttc gcccaggact ctagctatag ttctagtggt tggctacgta tactccggaa
     4861 tattaataga tcatggagat aattaaaatg ataaccatct cgcaaataaa taagtatttt
     4921 actgttttcg taacagtttt gtaataaaaa aacctataaa tattccggat tattcatacc
     4981 gtcccaccat cgggcgcgga tctcggtccg aaaccatgtc gtactaccat caccatcacc
     5041 atcacgatta cgatatccca acgaccgaaa acctgtattt tcagggcgcc atggatccaa
     5101 tgaatcaggt aacagactgg gttgacccat catttgatga ttttctagag tgtagtggcg
     5161 tctctactat tactgccaca tcattaggtg tgaataactc aagtcataga agaaaaaatg
     5221 ggccttctac attagaaagc agcagatttc cagcgagaaa aagaggaaat ctatcttcct
     5281 tagaacagat ttatggttta gaaaattcaa aagaatatct gtctgaaaat gaaccatggg
     5341 tggataaata taaaccagaa actcagcatg aacttgctgt gcataaaaag aaaattgaag
     5401 aagtcgaaac ctggttaaaa gctcaagttt tagaaaggca accaaaacag ggtggatcta
     5461 ttttattaat aacaggtcct cctggatgtg gaaagacaac gaccttaaaa atactatcaa
     5521 aggagcatgg tattcaagta caagagtgga ttaatccagt tttaccagac ttccaaaaag
     5581 atgatttcaa ggggatgttt aatactgaat caagcttcca tatgtttccc tatcagtctc
     5641 agatagcagt tttcaaagag tttctactaa gagcgacaaa gtataacaag ttacaaatgc
     5701 ttggagatga tctgagaact gataagaaga taattctggt tgaagattta cctaaccagt
     5761 tttatcggga ttctcatact ttacatgaag ttctaaggaa gtatgtgagg attggtcgat
     5821 gtcctcttat atttataatc tcggacagtc tcagtggaga taataatcaa aggttattgt
     5881 ttcccaaaga aattcaggaa gagtgttcta tctcaaatat tagtttcaac cctgtggcac
     5941 caacaattat gatgaaattt cttaatcgaa tagtgactat agaagctaac aagaatggag
     6001 gaaaaattac tgtccctgac aaaacttctc tagagttgct ctgtcaggga tgttctggtg
     6061 atatcagaag tgcaataaac agcctccagt tttcttcttc aaaaggagaa aacaacttac
     6121 ggccaaggaa aaaaggaatg tctttaaaat cagatgctgt gctgtcaaaa tcaaaacgaa
     6181 gaaaaaaacc tgatagggtt tttgaaaatc aagaggtcca agctattggt ggcaaagatg
     6241 tttctctgtt tctcttcaga gctttgggga aaattctata ttgtaaaaga gcatctttaa
     6301 cagaattaga ctcacctcgg ttgccctctc atttatcaga atatgaacgg gatacattac
     6361 ttgttgaacc tgaggaggta gtagaaatgt cacacatgcc tggagactta tttaatttat
     6421 atcttcacca aaactacata gatttcttca tggaaattga tgatattgtg agagccagtg
     6481 aatttctgag ttttgcagat atcctcagtg gtgactggaa tacacgctct ttactcaggg
     6541 aatatagcac atctatagct acgagaggtg tgatgcattc caacaaagcc cgaggatatg
     6601 ctcattgcca aggaggagga tcaagttttc gacccttgca caaacctcag tggtttctaa
     6661 taaataaaaa gtatcgggaa aattgcctgg cagcaaaagc actttttcct gacttctgcc
     6721 taccagcttt atgcctgcag actcagctat tgccatacct tgctctacta accattccaa
     6781 tgagaaatca agctcagatt tcttttatcc aagatattgg aaggctccct ctgaagcgac
     6841 actttggaag a
//
