LOCUS       Exported                7884 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pIRESneo-FLAG/HA Ago1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7884)
  AUTHORS   Meister G, Landthaler M, Patkaniowska A, Dorsett Y, Teng G, Tuschl T
  TITLE     Human Argonaute2 mediates RNA cleavage targeted by miRNAs and 
            siRNAs.
  JOURNAL   Mol Cell. 2004 Jul 23. 15(2):185-97.
  PUBMED    15260970
REFERENCE   2  (bases 1 to 7884)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7884
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      921..930
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             930..953
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             954..980
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          3640..3869
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3852..3871
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3927..4500
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4094..4111)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4321..4340
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4536..5339
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4599..4618)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5209..5228
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5371..5388)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5377..5601
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5648..5667
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5727..5743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5727..5743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5740..5762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5751..5767
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5775..5805)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5820..5841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5958..5975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6129..6717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6209..6228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6888..7748)
                     /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     7511..7530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7749..7853)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatcgcc gccgccatgg actacaagga cgacgatgac aagtaccctt
      961 atgacgtgcc cgattacgct agcggccgca tggaagcggg accctcggga gcagctgcgg
     1021 gcgcttacct gccccccctg cagcaggtgt tccaggcacc tcgccggcct ggcattggca
     1081 ctgtggggaa accaatcaag ctcctggcca attactttga ggtggacatc cctaagatcg
     1141 acgtgtacca ctacgaggtg gacatcaagc cggataagtg tccccgtaga gtcaaccggg
     1201 aagtggtgga atacatggtc cagcatttca agcctcagat ctttggtgat cgcaagcctg
     1261 tgtatgatgg aaagaagaac atttacactg tcacagcact gcccattggc aacgaacggg
     1321 tcgactttga ggtgacaatc cctggggaag ggaaggatcg aatctttaag gtctccatca
     1381 agtggctagc cattgtgagc tggcgaatgc tgcatgaggc cctggtcagc ggccagatcc
     1441 ctgttccctt ggagtctgtg caagccctgg atgtggccat gaggcacctg gcatccatga
     1501 ggtacacccc tgtgggccgc tccttcttct caccgcctga gggctactac cacccgctgg
     1561 ggggtgggcg cgaggtctgg ttcggctttc accagtctgt gcgccctgcc atgtggaaga
     1621 tgatgctcaa cattgatgtc tcagccactg ccttttataa ggcacagcca gtgattgagt
     1681 tcatgtgtga ggtgctggac atcaggaaca tagatgagca gcccaagccc ctcacggact
     1741 ctcagcgcgt tcgcttcacc aaggagatca agggcctgaa ggtggaagtc acccactgtg
     1801 gacagatgaa gaggaagtac cgcgtgtgta atgttacccg tcgccctgct agccatcaga
     1861 cattcccctt acagctggag agtggacaga ctgtggagtg cacagtggca cagtatttca
     1921 agcagaaata taaccttcag ctcaagtatc cccatctgcc ctgcctacaa gttggccagg
     1981 aacaaaagca tacctacctt cccctagagg tctgtaacat tgtggctggg cagcgctgta
     2041 ttaaaaagct gaccgacaac cagacctcga ccatgataaa ggccacagct agatccgctc
     2101 cagacagaca ggaggagatc agtcgcctga tgaagaatgc cagctacaac ttagatccct
     2161 acatccagga atttgggatc aaagtgaagg atgacatgac ggaggtgaca gggcgagtgc
     2221 tgccggcgcc catcttgcag tacggcggcc ggaaccgggc cattgccaca cccaatcagg
     2281 gtgtctggga catgcggggg aaacagttct acaatgggat tgagatcaaa gtctgggcca
     2341 tcgcctgctt cgcaccccaa aaacagtgtc gagaagaggt gctcaagaac ttcacagacc
     2401 agctgcggaa gatttccaag gatgcgggga tgcctatcca gggtcaacct tgtttctgca
     2461 aatatgcaca gggggcagac agcgtggagc ctatgttccg gcatctcaag aacacctact
     2521 cagggctgca gctcattatt gtcatcctgc cagggaagac gccggtgtat gctgaggtga
     2581 aacgtgtcgg agatacactc ttgggaatgg ctacgcagtg tgtgcaggtg aagaacgtgg
     2641 tcaagacctc acctcagact ctgtccaacc tctgcctcaa gatcaatgtc aaacttggtg
     2701 gcattaacaa catcctagtc ccacaccagc gctctgccgt ttttcaacag ccagtgatat
     2761 tcctgggagc agatgttaca caccccccag caggggatgg gaaaaaacct tctatcacag
     2821 cagtggtagg cagtatggat gcccacccca gccgatactg tgctactgtg cgggtacagc
     2881 gaccacggca agagatcatt gaagacttgt cctacatggt gcgtgagctc ctcatccaat
     2941 tctacaagtc cacccgtttc aagcctaccc gcatcatctt ctaccgagat ggggtgcctg
     3001 aaggccagct accccagata ctccactatg agctactggc cattcgtgat gcctgcatca
     3061 aactggaaaa ggactaccag cctgggatca cttatattgt ggtgcagaaa cgccatcaca
     3121 cccgcctttt ctgtgctgac aagaatgagc gaattgggaa gagtggtaac atcccagctg
     3181 ggaccacagt ggacaccaac atcacccacc catttgagtt tgacttctat ctgtgcagcc
     3241 acgcaggcat ccagggcacc agccgaccat cccattacta tgttctttgg gatgacaacc
     3301 gtttcacagc agatgagctc cagatcctga cgtaccagct gtgccacact tacgtacgat
     3361 gcacacgctc tgtctctatc ccagcacctg cctactatgc ccgcctggtg gctttccggg
     3421 cacgatacca cctggtggac aaggagcatg acagtggaga ggggagccac atatcggggc
     3481 agagcaatgg gcgggacccc caggccctgg ccaaagccgt gcaggttcac caggatactc
     3541 tgcgcaccat gtacttcgct tgagaattca gtggatccac tagtaacggc cgccagtgtg
     3601 ctggaattaa ttcgctgtct gcgagggcca gctgttgggg tgagtactcc ctctcaaaag
     3661 cgggcatgac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
     3721 cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct
     3781 ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg gccatacact tgagtgacaa
     3841 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac tgcaggtcga
     3901 gcatgcatct agggcggcca attccgcccc tctccctccc ccccccctaa cgttactggc
     3961 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgtgattttc caccatattg
     4021 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     4081 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     4141 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     4201 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     4261 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     4321 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     4381 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     4441 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     4501 gcttgccaca acccgggata attcctgcag ccaatatggg atcggccatt gaacaagatg
     4561 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4621 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4681 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4741 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4801 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4861 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4921 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4981 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5041 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg
     5101 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5161 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5221 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5281 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5341 gggatcaatt ctctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     5401 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     5461 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5521 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     5581 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgagtgcat
     5641 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5701 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5761 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5821 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5881 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5941 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6001 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6061 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6121 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6181 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6241 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6301 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     6361 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6421 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6481 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6541 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6601 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6661 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6721 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6781 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6841 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6901 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6961 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7021 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7081 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7141 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7201 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7261 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7321 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7381 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7441 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7501 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7561 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7621 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7681 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7741 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7801 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7861 ccccgaaaag tgccacctga cgtc
//
