LOCUS       Exported                7439 bp DNA     circular SYN 22-JUN-2020
DEFINITION  enables eukaryotic expression of N-terminally FLAG-HA-tagged fusion 
            proteins.
ACCESSION   .
VERSION     .
KEYWORDS    FLAG-HA-pcDNA3.1-
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7439)
  AUTHORS   Horn M, Geisen C, Cermak L, Becker B, Nakamura S, Klein C, Pagano M,
            Antebi A
  TITLE     DRE-1/FBXO11-Dependent Degradation of BLMP-1/BLIMP-1 Governs C. 
            elegans Developmental Timing and Maturation.
  JOURNAL   Dev Cell. 2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028.
  PUBMED    24613396
REFERENCE   2  (bases 1 to 7439)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2022 from SnapGene 6.0.5
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Cell. 
            2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028."
FEATURES             Location/Qualifiers
     source          1..7439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    245..471
                     /label=SB IR/DR(L)
                     /note="SB IR/DR(L)"
     misc_feature    538..793
                     /label=EFS-NS
                     /note="EFS-NS"
     misc_feature    771..1259
                     /label=NSP1-mut4-gBlock
     primer_bind     785..839
                     /label=A01-F1
     misc_feature    818..826
                     /label=Kozak Consensus Sequence
     CDS             827..1366
                     /codon_start=1
                     /label=nsp1
                     /translation="MESLVPGFNEKTHVQLSLPVLQVRDVLVRGFGDSVEEVLSEARQH
                     LKDGTCGLVEVEKGVLPQLEQPYVFIKRSDARTAPHGHVMVELVAELEGIQYGRSGETL
                     GVLVPHVGEIPVAYRKVLLRKNGSEGAGGHSYGADLKSFDLGDELGTDPYEDFQENWNT
                     KHSSGVTRELMRELNGG"
     CDS             1208..1213
                     /codon_start=1
                     /label=N128 K129 to S128 E129
                     /translation="SE"
     CDS             1385..1408
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1445..1468
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1472..1487
                     /label=3' Homology with pLVX-EF1a-IRES-Puro
     misc_feature    1473..1477
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1480..2053
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1541..1560)
                     /label=IRES-REV
     CDS             2086..2685
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2664..2723)
                     /label=Common-R1
     misc_feature    2692..2739
                     /label=Short PA
                     /note="Short PA"
     misc_feature    2746..2973
                     /label=SB IR/DR(R)
                     /note="SB IR/DR(R)"
     primer_bind     complement(3030..3047)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3036..3260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3306..3734
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3393..3412)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3603..3624
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3743..3763)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3748..4077
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3928..4063
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3990..4009
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4144..4938
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4198..4217)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4808..4827
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5112..5233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5149..5168)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5203..5222
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5282..5298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5282..5298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5295..5317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5306..5322
                     /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(5330..5360)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5375..5396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5513..5530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5684..6272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5764..5783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6443..7303)
                     /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     7066..7085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7304..7408)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttctagac
      241 tatacagttg aagtcggaag tttacataca cttaagttgg agtcattaaa actcgttttt
      301 caactactcc acaaatttct tgttaacaaa caatagtttt ggcaagtcag ttaggacatc
      361 tactttgtgc atgacacaag tcatttttcc aacaattgtt tacagacaga ttatttcact
      421 tataattcac tgtatcacaa ttccagtggg tcagaagttt acatacacta agttgactgt
      481 gcctttaaac agcttggaaa attccagaaa atgatgtcat ggctttagag aggatcctag
      541 gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc
      601 acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg
      661 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
      721 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
      781 gccagaacac aggaccggtg gtacccgtac ggccaccgcc gccaccatgg agagtcttgt
      841 acccggcttt aacgagaaaa cacatgttca attgtcattg cctgtgctgc aagtccggga
      901 cgtactggtt agggggttcg gagattcagt agaagaggtg ctgtctgagg cgaggcaaca
      961 tttgaaagac gggacgtgcg ggctggttga ggtggaaaag ggagtacttc cacagttgga
     1021 acaaccgtat gtttttatta agcggagtga tgccaggact gcgccgcacg gtcatgtcat
     1081 ggttgagctt gtcgcagagc ttgaaggcat tcaatatggg cgctccggtg aaactcttgg
     1141 agtacttgtg ccgcacgtag gcgaaattcc agtggcatat cggaaggtgc tgttgagaaa
     1201 aaatgggtct gagggcgcag ggggccacag ttatggagcc gacctcaaat cctttgacct
     1261 tggcgacgag ttggggacag atccctacga ggatttccag gaaaactgga atacgaaaca
     1321 tagcagtggc gttactcgcg agttgatgcg cgaactgaac ggcggcctcg aaggcggcgg
     1381 gggatggagc catccacaat tcgagaaagg cggtggttca ggaggaggta gcgggggtgg
     1441 atcatggtca catccgcagt ttgaaaagta aggatcccgc ccctctccct cccccccccc
     1501 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     1561 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     1621 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     1681 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     1741 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     1801 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     1861 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     1921 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     1981 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     2041 aacacgatga taagcttgcc acaacccaca aggagacgac cttccatgac cgagtacaag
     2101 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     2161 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     2221 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     2281 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     2341 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     2401 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     2461 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     2521 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     2581 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     2641 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctagctcga gaataaaaga
     2701 tctttatttt cattagatct gtgtgttggt tttttgtgtg aattcttgag tgtatgtaaa
     2761 cttctgaccc actgggaatg tgatgaaaga aataaaagct gaaatgaatc attctctcta
     2821 ctattattct gatatttcac attcttaaaa taaagtggtg atcctaactg acctaagaca
     2881 gggaattttt actaggatta aatgtcagga attgtgaaaa agtgagttta aatgtatttg
     2941 gctaaggtgt atgtaaactt ccgacttcaa ctgtataggc atgcggtaac cacgtgcgga
     3001 ccgaagctta agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3061 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3121 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3181 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3241 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3301 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3361 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3421 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3481 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3541 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3601 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3661 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3721 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3901 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3961 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4021 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4081 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4141 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4201 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4261 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4321 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4381 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4441 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4501 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4561 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4621 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4681 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4741 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4801 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4861 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4921 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4981 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5041 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5101 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5161 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5221 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5281 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5341 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5401 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5461 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5521 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5581 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5641 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5701 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5761 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5821 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5881 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5941 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6001 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6061 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6121 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6361 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6421 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6481 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6541 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6601 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6661 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6721 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6781 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6841 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6901 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6961 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7021 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7081 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7141 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7201 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7261 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7321 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7381 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
