LOCUS       pcDNA_Zeo_HA_DD-N_mScarlet-I_QtSig        6123 bp    DNA     circular UNA 10-OCT-2019
DEFINITION  .
ACCESSION   urn.local...w-ixztrrm
VERSION     urn.local...w-ixztrrm
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     940..966
                     /Tm=67
                     /Sequence="ACCATGGTTTCTAAAGGCGAGGCCGTG"
                     /Mismatch_Positions="A===TG==TTCTA==GG==AGG==G=="
                     /Mismatches=14
                     /Extension="TCCAGTGTGGTGGAATTCGCC"
                     /Description="<html><body></body></html>"
                     /label="mScarlet_overlaps_iBOX_for"
     primer_bind     complement(941..963)
                     /Tm=65
                     /Sequence="GGTGGCGAATTCCACCACACTGG"
                     /Description="<html><body></body></html>"
                     /label="pcDNA_Zeo_lin_rev"
     primer_bind     947..969
                     /Tm=62
                     /Sequence="TGGTGGAATTCGCCACCATGTAC"
                     /Description="<html><body></body></html>"
                     /label="mScarlet-I-_EcoRI_for"
     CDS             964..966
                     /codon_start=1
                     /product="start codon"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
     CDS             964..2085
                     /codon_start=1
                     /transl_table=1
                     /translation="MYPYDVPDYAGTGVQVETISPGDGRTFPKRGQTCVVHYTGMLED
                     GKKVDSSRDRNKPFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGI
                     IPPHATLVFDVELLKPEGSGVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERV
                     MNFEDGGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPED
                     GVLKGDIKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVE
                     QYERSEGRHSTGGMDELYKGGGGSGGGGSKGFTVGSLIQ*"
                     /label="Feature 28"
     CDS             967..993
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1000..1320
                     /codon_start=1
                     /note="L106P mutant of FKBP12"
                     /product="destabilization domain that can be stabilized by
                     Shield1 in the ProteoTuner™ system"
                     /transl_table=1
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVE
                     LLKPE"
                     /label="DD-N"
     CDS             1321..1329
                     /codon_start=1
                     /transl_table=1
                     /translation="GSG"
                     /label="Linker"
     primer_bind     1321..1342
                     /Tm=59
                     /Sequence="GGATCTGGCGTTTCTAAAGGCG"
                     /Extension="CTTGTCGTCGTCGTCCTTGTAGTC"
                     /Description="<html><body></body></html>"
                     /label="GSG_mScarlet_overlaps_Zip14_for"
     primer_bind     1327..1350
                     /Tm=61
                     /Sequence="GGAGTTTCTAAAGGCGAGGCCGTG"
                     /Mismatch_Positions="==A====================="
                     /Mismatches=1
                     /Extension="ATCCGGCGGAGGTGGAAGC"
                     /Description="<html><body>  </body></html>"
                     /label="mScarlet_overlaps_NB_for"
     CDS             1330..2022
                     /codon_start=1
                     /note="/Transferred_From=pCAG-Avilabel3.1"
                     /note="/Transferred_Similarity=90.62%"
                     /note="/created_by=jeffe"
                     /note="/modified_by=jeffe"
                     /transl_table=1
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDG
                     GAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGD
                     IKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSE
                     GRHSTGGMDELYK"
                     /label="mScarlet-I"
     variation       1546..1548
                     /note="/Transferred_From=pCAG-Avilabel3.1"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=jeffe"
                     /note="/modified_by=jeffe"
                     /label="T74I results in mScarlet-I (fast maturation)"
     primer_bind     complement(2000..2096)
                     /Tm=85
                     /Sequence="CGAGCGGCCGCTCATTGGATCAGGCTGCCCACGGTGAAGCCCTTAGA
                     TCCGCCTCCGCCAGATCCTCCGCCGCCCTTGTACAGCTCGTCCATTCCGC"
                     /Description="<html><body>  </body></html>"
                     /label="mScarlet-I_QtSig_NotI_rev"
     primer_bind     complement(2000..2022)
                     /Tm=62
                     /Sequence="CTTGTACAGCTCGTCCATTCCGC"
                     /Description="<html><body></body></html>"
                     /label="mScarlet_GA_rev"
     primer_bind     complement(2000..2022)
                     /Tm=62
                     /Sequence="CTTGTACAGCTCGTCCATTCCGC"
                     /Extension="TCA"
                     /Description="<html><body></body></html>"
                     /label="mScalet_overlaps_NB_rev"
     primer_bind     complement(2000..2042)
                     /Tm=66
                     /Sequence="GCTTCCAGGCCGCCAGATCCCTTGTACAGCTCGTCCATTCCGC"
                     /Mismatch_Positions="G=TT=CAGG==G==AGAT===================
                     ======"
                     /Mismatches=12
                     /Extension="GTCCAATCCTCG"
                     /Description="<html><body></body></html>"
                     /label="mScarlet_overlaps_iBOX_rev"
     misc_feature    2015..2020
                     /note="/Transferred_From=pCAG-Avilabel3.1"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=jeffe"
                     /label="BsrGI"
     CDS             2023..2052
                     /codon_start=1
                     /transl_table=1
                     /translation="GGGGSGGGGS"
                     /label="Linker"
     primer_bind     complement(2032..2102)
                     /Tm=81
                     /Sequence="TAGACTCGAGCGGCCGCTCATTGGATCAGGCTGCCCACGGTGAAGCC
                     CTTAGATCCGCCTCCGCCAGATCC"
                     /Description="<html><body></body></html>"
                     /label="DD-mEos4b_GeoTag_rev"
     CDS             2053..2082
                     /codon_start=1
                     /transl_table=1
                     /translation="KGFTVGSLIQ"
                     /label="GeoTag"
     primer_bind     complement(2064..2093)
                     /Tm=55
                     /Sequence="GCGCCAGAGCCTTGGATCAGGCTGCCCACG"
                     /Mismatch_Positions="===C=A=AG=C==================="
                     /Mismatches=5
                     /Extension="GCTGAAGTTTGTG"
                     /Description="<html><body></body></html>"
                     /label="DD-Eos_GeoSig_GA_rev"
     primer_bind     complement(2067..2085)
                     /Tm=54
                     /Sequence="TCATTGGATCAGGCTGCCC"
                     /Extension="GCTCCCTTAGCTCCCTGAGACG"
                     /Description="<html><body></body></html>"
                     /label="mEos4b_BM40_GA_rev"
     primer_bind     complement(2083..2108)
                     /Tm=71
                     /Sequence="TCACTTGTACAGCTCGTCCATTCCGC"
                     /Mismatch_Positions="T=A==TGTACAG=TC=TCCATT=CGC"
                     /Mismatches=20
                     /Extension="GCCCTCTAGACTCGAGCGGCCGC"
                     /Description="<html><body></body></html>"
                     /label="mScarlet_overlaps_Zip14_rev"
     misc_feature    2086..2106
                     /label="terminal"
     primer_bind     2086..2105
                     /Tm=63
                     /Sequence="GCGGCCGCTCGAGTCTAGAG"
                     /Description="<html><body></body></html>"
                     /label="pcDNA_Zeo_lin_for"
     polyA_signal    2136..2360
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2406..2834
                     /snapgene_direction="RIGHT"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2848..3178
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     rep_origin      3029..3164
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        3226..3273
                     /note="synthetic bacterial promoter&nbsp;"
                     /label="EM7 promoter"
     CDS             3292..3666
                     /codon_start=1
                     /gene="<i>Sh&nbsp;ble</i> from&nbsp;<i>Streptoalloteichus
                     hindustanus</i>"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin™"
                     /product="antibiotic-binding protein"
                     /transl_table=1
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD
                     DVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPW
                     GREFALRDPAGNCVHFVAEEQD*"
                     /label="BleoR"
     polyA_signal    3796..3917
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3966..3982)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3990..4006
                     /bound_moiety="<i>lac</i> repressor encoded by
                     <i>lacI</i>"
                     /note="The <i>lac</i> repressor binds to the <i>lac</i>
                     operator to inhibit transcription in <i>E. coli</i>. This
                     inhibition can be relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        join(complement(4039..4044),complement(4021..4038),
                     complement(4014..4020))
                     /note="promoter for the <i>E. coli lac</i> operon"
                     /label="lac promoter"
     protein_bind    4059..4080
                     /bound_moiety="<i>E. coli</i> catabolite activator
                     protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4368..4956)
                     /snapgene_direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             join(complement(5919..5987),complement(5127..5918))
                     /codon_start=1
                     /gene="<i>bla</i>"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="AmpR"
     promoter        complement(5988..6092)
                     /gene="<i>bla</i>"
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgtacc catacgatgt tccagattac gctggtaccg gagtgcaggt ggaaaccatc
     1021 tccccaggag acgggcgcac cttccccaag cgcggccaga cctgtgtggt gcactacacc
     1081 gggatgcttg aagatggaaa gaaagtcgat tcctcccggg acagaaacaa gccctttaag
     1141 tttatgctag gcaagcagga ggtgatccga ggctgggaag aaggggttgc ccagatgagt
     1201 gtgggtcaga gagccaaact gactatatct ccagattatg cctatggtgc cactgggcac
     1261 ccaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct aaaaccggaa
     1321 ggatctggcg tttctaaagg cgaggccgtg atcaaagagt tcatgaggtt caaggtgcac
     1381 atggaaggca gcatgaacgg ccacgagttc gagatcgaag gcgaaggcga gggcagacct
     1441 tacgagggaa ctcagaccgc caagctgaaa gtgaccaaag gcggccctct gcctttcagc
     1501 tgggacatcc tgtctccaca gtttatgtac ggcagcaggg ccttcatcaa gcaccccgct
     1561 gacatccccg actactacaa gcagtctttc ccagagggct tcaagtggga gagagtgatg
     1621 aacttcgagg acggcggagc cgtgactgtg acacaggaca catctctgga agatgggacc
     1681 ctgatctaca aagtgaagct gcggggcaca aacttccctc cagacggccc tgtgatgcag
     1741 aaaaagacaa tgggctggga agccagcacc gagagactgt accctgaaga tggcgtgctg
     1801 aagggcgaca tcaagatggc cctgagactg aaggatggcg gcagatacct ggccgacttc
     1861 aagaccacct acaaggccaa gaaacctgtg cagatgcctg gcgcctacaa cgtggacaga
     1921 aagctggaca tcaccagcca caacgaggac tacaccgtgg tggaacagta cgagagaagc
     1981 gaggggagac acagcacagg cggaatggac gagctgtaca agggcggcgg aggatctggc
     2041 ggaggcggat ctaagggctt caccgtgggc agcctgatcc aatgagcggc cgctcgagtc
     2101 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2161 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2221 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2281 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2341 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2401 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2461 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2521 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     2581 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2641 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2701 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2761 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2821 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2881 tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2941 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3001 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3061 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3121 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3181 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
     3241 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
     3301 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
     3361 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
     3421 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
     3481 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
     3541 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
     3601 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
     3661 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3721 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3781 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3841 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3901 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     3961 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4021 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4081 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4141 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4201 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4261 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4321 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4381 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4441 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4501 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4561 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4621 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4681 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4741 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4801 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4861 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4921 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4981 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5041 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5101 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5161 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5221 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5281 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5341 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5401 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5461 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5521 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5581 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5641 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5701 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5761 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5821 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5881 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5941 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6001 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6061 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6121 gtc
//
