LOCUS       sequence-176430-        7078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Giardia Dicer in Sf9 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7078)
  AUTHORS   MacRae IJ, Ma E, Zhou M, Robinson CV, Doudna JA
  TITLE     In vitro reconstitution of the human RISC-loading complex.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Jan 15;105(2):512-7. doi: 
            10.1073/pnas.0710869105. Epub 2008 Jan 4.
  PUBMED    18178619
REFERENCE   2  (bases 1 to 7078)
  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.0710869105"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2008-01-15- 15"; volume: "105"; issue: "2"; pages: "512-7"
COMMENT     Sequence Label: pFastBac-Giardia Dicer
FEATURES             Location/Qualifiers
     source          1..7078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        179..270
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     195..212
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             337..354
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             376..396
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2862..2996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2899..2918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2953..2972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3025..3190
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3374..3828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3461..3480)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3670..3691
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3854..3958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3959..4027,4028..4819)
                     /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(4177..4196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4990..5578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5479..5498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5732..5749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5883..6107
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6174..6707)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6877..6882,6883..6899,6900..6905))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg ccggtggtgc
       61 tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg
      121 cccaggactc tagctatagt tctagtggtt ggctacgtat actccggaat attaatagat
      181 catggagata attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt
      241 aacagttttg taataaaaaa acctataaat attccggatt attcataccg tcccaccatc
      301 gggcgcggat ctcggtccga aaccatgtcg tactaccatc accatcacca tcacgattac
      361 gatatcccaa cgaccgaaaa cctgtatttt cagggcgcca tgcatgcttt gggacactgt
      421 tgcacagttg tgactactag aggaccatcc cactggttgc tacttctaga cactcacctg
      481 ggcaccttgc cagggtttaa ggttagtgca ggccgagggc ttcccgccgc agaggtgtac
      541 tttgaagcgg gtccgagggt gtctctctct cgaactgatg caactatagt agccgtgtat
      601 cagtccattc tctttcagct gctgggaccc acatttcctg cttcatggac tgagattgga
      661 gcaacaatgc ctcacaatga atacactttc cctcgattta tatccaatcc accacaattc
      721 gccaccctgg catttttacc cttactatct cctaccagcc ctctggactt gcgtgcatta
      781 atggtcactg cacaactcat gtgtgatgca aagcgcttgt cagatgaata tacagactat
      841 tccactttat ctgcatccct ccatgggcgt atggttgcaa ctcccgaaat aagctggtct
      901 ctttatgtcg ttcttgggat cgattctacc caaactagcc tttcttactt taccagagca
      961 aatgaatcaa taacatacat gagatactat gcaacagccc acaatattca cctgcgtgct
     1021 gcagatcttc cgcttgtggc agcagtcaga ttagacgatc taaaagacca ccagattccc
     1081 gcgcctggat cctgggatga tttggctccc aagcttcgct tcctgccgcc tgagctctgc
     1141 ctactgctgc cagatgaatt tgatctaatc agggtccagg cgcttcaatt tctaccagag
     1201 attgctaagc acatatgtga catacagaat acaatctgtg ccctggataa aagctttcct
     1261 gactgtgggc ggatcggtgg cgagcgatac tttgcaatca ctgccggact tcggctcgat
     1321 caggggcgtg gacgagggct tgccggttgg agaacaccct ttgggccttt tggtgtaagt
     1381 cacaccgatg ttttccagcg actcgaattg ctaggagatg ctgtgttagg ctttatcgtg
     1441 actgcccgcc tcctttgcct ttttccagat gcgtctgtgg gaacacttgt tgagctaaag
     1501 atggagcttg ttcgcaatga ggctctaaac tatcttgtac aaacgcttgg acttcctcag
     1561 ttggcggagt tttccaacaa ccttgtggcg aagagcaaaa catgggcaga tatgtatgag
     1621 gagatcgttg gatcaatctt tacgggacct aatggaatct atggctgtga ggaatttctt
     1681 gcgaagacgc ttatgagtcc cgaacactcc aagacagtag gatctgcctg tccagatgca
     1741 gtcaccaagg catcaaagcg tgtttgcatg ggagaagcgg gggcgcatga attcagaagc
     1801 cttgtggact atgcttgtga gcaaggcatt agtgtcttct gttcttcgcg ggtgtcaact
     1861 atgtttctcg agcgtctcag agacattcca gcagaggaca tgctagattg gtaccgactt
     1921 ggtatccagt tttcgcatcg ttcaggccta tcaggacctg gcggcgtcgt atcagttata
     1981 gacataatga cacatttggc tcgaggccta tggctgggct ctccaggctt ctatgttgaa
     2041 cagcaaactg ataagaatga gtcggcttgt ccgcccacta tacctgtttt atatatctat
     2101 catcgctctg tgcagtgtcc tgttttatat gggtcgctca cagaaacccc tacagggccc
     2161 gttgcttcta aggttctcgc tctctatgag aagattctgg catatgagtc atcaggaggt
     2221 agtaagcata tagcagctca gacagttagc agatctctgg ccgtacccat tcctagtggc
     2281 actatcccct tcctgattcg gttattgcaa atagcactaa ctcctcacgt gtaccaaaaa
     2341 cttgagcttc ttggagacgc attcctgaag tgcagccttg ctctccatct ccacgctctc
     2401 caccccacgc tcacagaggg cgctcttaca cgcatgcggc aatctgcaga aacaaattct
     2461 gtactgggaa gattgacaaa aaggtttcct tctgtagttt ctgaggttat tatagaatcc
     2521 catccgaaaa tacagcctga cagcaaggtt tatggcgata catttgaagc cattttggca
     2581 gcaattcttc ttgcgtgcgg ggaagaggca gcaggtgctt ttgttcgaga gcatgttctc
     2641 ccacaagtag tagctgatgc gtaggcggcc gcgtcgacac tagtgcggcc gctttcgaat
     2701 ctagagcctg cagtctcgag gcatgcggta ccaagcttgt cgagaagtac tagaggatca
     2761 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2821 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2881 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2941 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat
     3001 cactgcttga gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg
     3061 tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac
     3121 tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc aactattttg
     3181 tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact
     3241 ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct
     3301 gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg
     3361 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3421 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3481 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3541 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3601 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3661 ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3721 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3781 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     3841 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3901 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3961 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4021 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4081 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4141 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4201 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4261 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4321 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4381 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4441 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4501 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4561 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4621 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4681 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4741 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4801 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4861 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4921 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4981 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5041 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5101 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5161 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5221 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5281 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5341 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5401 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5461 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5521 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5581 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5641 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5701 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5761 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca
     5821 gccgcgtaac ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg
     5881 ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa
     5941 gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc
     6001 atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg
     6061 tcgtattaaa gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac
     6121 aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg
     6181 gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag
     6241 agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc
     6301 gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc
     6361 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc
     6421 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg
     6481 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag
     6541 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
     6601 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
     6661 ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     6721 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
     6781 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
     6841 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
     6901 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
     6961 agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc
     7021 acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacgg
//
