LOCUS       Exported                7000 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  encodes multiple copies of an epitope tag HA.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG_smFP HA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7000)
  AUTHORS   Viswanathan S, Williams ME, Bloss EB, Stasevich TJ, Speer CM, Nern 
            A, Pfeiffer BD, Hooks BM, Li WP, English BP, Tian T, Henry GL, 
            Macklin JJ, Patel R, Gerfen CR, Zhuang X, Wang Y, Rubin GM, Looger 
            LL
  TITLE     High-performance probes for light and electron microscopy.
  JOURNAL   Nat Methods. 2015 Jun;12(6):568-76. doi: 10.1038/nmeth.3365. Epub 
            2015 Apr 27.
  PUBMED    25915120
REFERENCE   2  (bases 1 to 7000)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7000
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     intron          216..1232
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1156..1178
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1240..1259
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1358..1384
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1388..1414
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1418..1444
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1973..1999
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2006..2032
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2045..2071
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2078..2104
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2315..2341
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2345..2371
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2375..2401
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2450..2582
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2565..2584
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2591..3179
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2644..2664)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3062..3073)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3206..3223)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3212..3323
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3419..3439)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3419..3437)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3458..3474)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3576..3595)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3641..3696
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3695..3714)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4057..4073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4081..4097
                     /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(4105..4135)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4150..4171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4230..4425
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4276..4411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4338..4357
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4431..4565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4468..4487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4522..4541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4633..4650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4804..5392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4884..4903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5563..6423)
                     /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     6186..6205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6424..6528)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6559..6938
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6940..215
                     /label=chicken beta-actin promoter
ORIGIN
        1 tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggs
       61 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
      121 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
      181 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
      241 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      301 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
      361 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
      421 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
      481 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
      541 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
      601 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
      661 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
      721 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
      781 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
      841 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
      901 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
      961 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     1021 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     1081 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     1141 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     1201 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     1261 tgtctcatca ttttggcaaa gaattcggta ccgcgggccc ggcgcgccag ctggatatct
     1321 ccggacccgg gctagcgcaa ttgcggccgc caccatgtac ccttatgatg tgcccgatta
     1381 tgctggctac ccttatgatg ttcctgatta cgccggatat ccgtatgacg tgccagacta
     1441 cgcgggaggt gtgagcaagg gagaggagtt gtttaccggc gtggtcccca tactggtgga
     1501 gttggatggc gacgttaatg gacataaatt ctcggtccgc ggcgagggag agggagacgc
     1561 caccaatggc aagctgacgc ttaagttcat ttgcacgact ggaaaattgc ctgtcccctg
     1621 gccgaccctg gtcaccacac tgggcggagg agtgcagtgc ttctcccgtt acccagacca
     1681 catgaagcag cacgatttct tcaagagcgc aatgccggag ggatacgtgc aagaacggac
     1741 tatttccttc aaggatgacg gaacctataa gacccgtgcg gaggtcaaat tcgagggtga
     1801 caccctggtg aaccgaattg aactcaaagg aatcgatttc aaggaggatg gaaatattct
     1861 gggtcacaag ctggaataca acttcaacag ccataatgtg tacattacgg ctgataagca
     1921 gaagaacggc attaaggcca atttcaagat ccgccacaac gttgagggtg gatatcccta
     1981 cgacgtgccc gattatgcgg gcggatatcc gtatgatgtt ccagattatg ctggtggtgg
     2041 aggctatccc tatgatgtcc ccgactacgc cggaggttac ccatacgacg tgcccgacta
     2101 cgctggaggc gatggcagcg tgcagctggc agatcattat caacagaata cccccatagg
     2161 tgatggcccc gttctgcttc cagataatca ctacctttcc acccagagcg tgctttcgaa
     2221 agacccgaac gaaaagcgtg atcacatggt cctgctggag tttgtgaccg cggctggaat
     2281 caccctgggt atggacgaac tctacaaggg tggctacccc tacgatgtgc cggattacgc
     2341 tggatatccg tatgacgtac cggactatgc cggttacccg tacgatgtcc cggactacgc
     2401 ttaaaccggt gctcgaggta ccgatgaata gctaaggtcg aggccgcagg taagtatcaa
     2461 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     2521 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     2581 aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2641 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2701 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2761 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2821 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2881 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2941 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3001 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3061 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3121 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3181 tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3241 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3301 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3361 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc
     3421 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3481 attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     3541 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     3601 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     3661 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     3721 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     3781 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     3841 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     3901 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     3961 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     4021 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4081 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4141 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4201 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     4261 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4321 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4381 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     4441 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4501 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4561 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6421 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6481 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6541 aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg
     6601 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6661 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6721 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6781 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6841 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6901 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
     6961 gcttcactct ccccatctcc cccccctccc cacccccaat
//
