LOCUS       sequence-456183-        5611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hHus1 in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5611)
  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 5611)
  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-hHus1
FEATURES             Location/Qualifiers
     source          1..5611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  18..183
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      367..821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(454..473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     663..684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        847..951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(952..1020,1021..1812)
                     /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(1170..1189)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1983..2571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2472..2491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2725..2742
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2876..3100
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3167..3700)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3870..3875,3876..3892,3893..3898))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        4250..4341
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4266..4283
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5271..5294
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    5466..5600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5503..5522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5557..5576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tgagcctagg agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt
       61 taattttcgt attagcttac gacgctacac ccagttccca tctattttgt cactcttccc
      121 taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc
      181 acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg
      241 acaaaccgtc atcttcggct actttttctc tgtcacagaa tgaaaatttt tctgtcatct
      301 cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttgcagc ctgaatggcg
      361 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttaa
      661 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
      721 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
      781 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
      841 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      901 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      961 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1021 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1081 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1141 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1201 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1261 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1321 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1381 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     1441 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     1501 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     1561 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     1621 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     1681 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1741 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1801 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     1861 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1921 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1981 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2041 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2101 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2161 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2221 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2281 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2341 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2401 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2461 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2521 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2581 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2641 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     2701 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     2761 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga ccagccgcgt
     2821 aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg
     2881 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa
     2941 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg
     3001 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt
     3061 aaagaggggc gtggccaagg gcatggtaaa gactatattc gcggcgttgt gacaatttac
     3121 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     3181 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     3241 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     3301 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     3361 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     3421 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     3481 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     3541 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     3601 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     3661 gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt
     3721 aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag
     3781 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
     3841 tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact
     3901 gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg
     3961 aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc
     4021 gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct
     4081 ggcttcagga gatcggaaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg
     4141 atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccaggact
     4201 ctagctatag ttctagtggt tggctacgta tactccggaa tattaataga tcatggagat
     4261 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     4321 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
     4381 tcccggtccg aagcgcgcgg aattcaaagg cctacgtcga caccatgaag tttcgggcca
     4441 agatcgtgga cggggcctgt ctgaaccact tcacacgaat cagtaacatg atagccaagc
     4501 ttgccaaaac ctgcaccctc cgcatcagcc ctgataagct taacttcatc ctttgtgaca
     4561 agctggctaa tggaggagtg agcatgtggt gtgagctgga acaggagaac ttcttcaacg
     4621 aatttcaaat ggagggtgtc tctgcagaaa acaatgagat ttatttagag ctaacatcgg
     4681 aaaacttatc tcgagccttg aagactgccc agaatgccag ggctttgaaa atcaaactga
     4741 ctaataaaca ctttccctgc ctcacggtct ccgtggagct gttatctatg tcaagcagta
     4801 gccgcattgt gacccatgac atccccataa aggtgattcc taggaaattg tggaaggact
     4861 tacaagaacc ggtggtccca gatcctgatg ttagtattta tttaccagtc ttgaagacta
     4921 tgaagagtgt tgtggaaaaa atgaaaaaca tcagcaatca ccttgttatt gaagcaaacc
     4981 tagatggaga attgaatttg aaaatagaaa ctgaattagt atgtgttaca actcatttta
     5041 aagatcttgg aaatcctcca ttagcctctg aaagcaccca tgaggacaga aacgtggaac
     5101 acatggctga agtgcacata gatattagga agctcctaca gtttcttgct ggacaacaag
     5161 taaatcccac aaaggcctta tgcaatattg tgaataacaa gatggtgcat tttgatctgc
     5221 ttcatgaaga cgtgtccctt cagtatttca tccctgcgct gtccgccatg gattacaaag
     5281 acgatgacga caaggctagc tagtctagag cctgcagtct cgaggcatgc ggtaccaagc
     5341 ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag gttttacttg
     5401 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     5461 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5521 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5581 tatcttatca tgtctggatc tgatcactgc t
//
