LOCUS       Exported                7450 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 7450)
  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 7450)
  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..7450
                     /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    775..1270
                     /label=NSP1-mutant1gblock
     primer_bind     785..839
                     /label=A01-F1
     misc_feature    818..826
                     /label=Kozak Consensus Sequence
     CDS             827..1377
                     /codon_start=1
                     /label=nsp1
                     /translation="MESLVPGFNEKT*VSDMFNCHCLCCKSGTYWLGGSEIQ*KRCCLR
                     RGNI*KTGRAGWLRWKREYFHSWNNRMFLLSGVMPGLRRTVMSWLSLSQSLKAFNMGAP
                     VKLLEYLCRT*AKFQWHIGRCC*EKMGTRAQGATVMEPTSNPLTLATSWGQIPTRISRK
                     TGIRNIAVALLAS*CAN*TA"
     misc_feature    863..873
                     /label=Triple stops
                     /note="Triple stops"
     CDS             1396..1419
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1456..1479
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1483..1498
                     /label=3' Homology with pLVX-EF1a-IRES-Puro
     misc_feature    1484..1488
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1491..2064
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1552..1571)
                     /label=IRES-REV
     CDS             2097..2696
                     /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(2675..2734)
                     /label=Common-R1
     misc_feature    2703..2750
                     /label=Short PA
                     /note="Short PA"
     misc_feature    2757..2984
                     /label=SB IR/DR(R)
                     /note="SB IR/DR(R)"
     primer_bind     complement(3041..3058)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3047..3271
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3317..3745
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3404..3423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3614..3635
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3754..3774)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3759..4088
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3939..4074
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4001..4020
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4155..4949
                     /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(4209..4228)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4819..4838
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5123..5244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5160..5179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5214..5233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5293..5309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5293..5309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5306..5328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5317..5333
                     /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(5341..5371)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5386..5407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5524..5541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5695..6283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5775..5794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6454..7314)
                     /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     7077..7096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7315..7419)
                     /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 cataagtaag tgacatgttc aattgtcatt gcctgtgctg
      901 caagtccggg acgtactggt tagggggttc ggagattcag tagaagaggt gctgtctgag
      961 gcgaggcaac atttgaaaga cgggacgtgc gggctggttg aggtggaaaa gggagtactt
     1021 ccacagttgg aacaaccgta tgtttttatt aagcggagtg atgccaggac tgcgccgcac
     1081 ggtcatgtca tggttgagct tgtcgcagag cttgaaggca ttcaatatgg gcgctccggt
     1141 gaaactcttg gagtacttgt gccgcacgta ggcgaaattc cagtggcata tcggaaggtg
     1201 ctgttgagaa aaaatgggaa caagggcgca gggggccaca gttatggagc cgacctcaaa
     1261 tcctttgacc ttggcgacga gttggggaca gatccctacg aggatttcca ggaaaactgg
     1321 aatacgaaac atagcagtgg cgttactcgc gagttgatgc gcgaactgaa cggcggcctc
     1381 gaaggcggcg ggggatggag ccatccacaa ttcgagaaag gcggtggttc aggaggaggt
     1441 agcgggggtg gatcatggtc acatccgcag tttgaaaagt aaggatcccg cccctctccc
     1501 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     1561 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     1621 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     1681 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     1741 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     1801 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     1861 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     1921 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     1981 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2041 ttcctttgaa aaacacgatg ataagcttgc cacaacccac aaggagacga ccttccatga
     2101 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     2161 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     2221 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     2281 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     2341 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     2401 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     2461 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     2521 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     2581 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     2641 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctagctcg
     2701 agaataaaag atctttattt tcattagatc tgtgtgttgg ttttttgtgt gaattcttga
     2761 gtgtatgtaa acttctgacc cactgggaat gtgatgaaag aaataaaagc tgaaatgaat
     2821 cattctctct actattattc tgatatttca cattcttaaa ataaagtggt gatcctaact
     2881 gacctaagac agggaatttt tactaggatt aaatgtcagg aattgtgaaa aagtgagttt
     2941 aaatgtattt ggctaaggtg tatgtaaact tccgacttca actgtatagg catgcggtaa
     3001 ccacgtgcgg accgaagctt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     3061 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3121 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3181 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3241 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3301 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3361 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3421 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3481 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3541 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3601 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3661 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3721 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3781 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3841 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3901 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3961 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4021 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4081 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4141 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4201 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4261 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4321 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4381 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4441 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4501 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4561 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4621 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4681 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4741 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4801 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4861 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4921 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4981 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5041 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5101 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5221 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5281 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5341 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5401 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5461 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5521 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5581 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7381 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7441 acctgacgtc
//
