LOCUS       Exported                6769 bp ds-DNA     circular SYN 15-JUN-2022
DEFINITION  A fusion protein of the SunTag10xGCN4 v4 peptide array to a 
            mitochondrial targeting domain and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA4TO-mito-mCherry-10xGCN4_v4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6769)
  AUTHORS   Tanenbaum ME, Gilbert LA, Qi LS, Weissman JS, Vale RD
  TITLE     A Protein-Tagging System for Signal Amplification in Gene Expression
            and Fluorescence Imaging.
  JOURNAL   Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi: 
            10.1016/j.cell.2014.09.039.
  PUBMED    25307933
REFERENCE   2  (bases 1 to 6769)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6769
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(81..410)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(149..168)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     395..415
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(424..852)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(534..555)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     746..765
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    898..1122
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1111..1128
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1212..1268)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1284..1340)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1356..1412)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1428..1484)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1500..1556)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1572..1628)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1644..1700)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1716..1772)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1788..1844)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1860..1916)
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(1989..2693)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2264..2283)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2533..2551
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3012..3036)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3046..3064
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3067..3085
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3087..3290)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3116..3136)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3291..3670
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3842..3861
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3936..4040
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4041..4901
                     /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     complement(4259..4278)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5072..5660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5561..5580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5814..5831
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5948..5969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5984..6014
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6022..6038
                     /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)."
     primer_bind     6027..6049
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6046..6062
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6046..6062
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6111..6232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6122..6141)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6176..6195
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6362..6736)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(6755..33)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
ORIGIN
        1 gccgatatac tatgccgatg attaattgtc aacacgtgct gatcagatcc gaaaatggat
       61 atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
      121 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
      181 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
      241 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
      301 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
      361 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
      421 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
      481 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
      541 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
      601 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
      661 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
      721 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
      781 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
      841 tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca
      901 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
      961 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     1021 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     1081 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggg
     1141 tttaaacggg ccctctagac tcgagtatta tgatcgatcc atatgtttac cgcggtgcct
     1201 gagcctgagc cctttttgag cctagcaact tcgttctcaa gatgataatt tttcgatagc
     1261 aattcctcgc cagaaccact cccctttttt agtctcgcta cctcattttc cagatgataa
     1321 tttttactca gtaattcttc acccgaccct gaacctttct ttaatcgagc tacttcgttt
     1381 tcgaggtggt aattctttga aagtagttcc tcaccactgc ccgatccctt ttttaatcga
     1441 gcgacctcat tttcaagatg atagttcttc gagagcagtt cctctcctga gccggaaccc
     1501 ttcttcaaac gcgctacctc gttctcaaga tgataattct tggatagtag ctcttcacca
     1561 ctgccacttc ccttttttaa gcgtgccacc tcattttcga ggtggtagtt tttagataat
     1621 agctcttctc cagaaccact gccctttttt agccgagcca cttcgttttc gagatgataa
     1681 tttttcgaga gtaactcctc acccgatccg gaccctttct ttagtcgtgc tacttcgttc
     1741 tccaaatgat aatttttgct cagcaactcc tccccacttc cgctcccctt tttcagtcta
     1801 gctacctcat tttccaggtg gtaattcttt gaaagcagtt cttctccact gccagaacct
     1861 ttcttaagac gagccacttc gttctcaaga tgataattct tgctcaaaag ttcttccgga
     1921 ccgctttgta caaaactagt catggtggca cgcgtgatcg atcgatctgc ggccgcgcca
     1981 ccaccaccct tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
     2041 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
     2101 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
     2161 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     2221 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     2281 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     2341 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     2401 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     2461 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     2521 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     2581 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     2641 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct cacaccgaat
     2701 tcggcgaccg gtaagcttcc agcatgtact atcttggggt tgtctttctg gatgtgaagg
     2761 tttatcatag ctttatttcg atgatcttta acataaaatc ttttgtaagc tagataacca
     2821 attgcagctg tcccagcagc aatggtaact gctgcgatcc attcaactcg tacgctggaa
     2881 ctggaagtca gactcatggt ggcggatccg agctcggtac caagcttaag tttaaacgct
     2941 agagtccgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca gcgtggatgg
     3001 cgtctccagg cgatctgacg gttcactaaa cgagctcgtc gacgatctct atcactgata
     3061 gggagatctc tatcactgat agggagagct ctgcttatat agacctccca ccgtacacgc
     3121 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     3181 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     3241 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     3301 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     3361 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     3421 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     3481 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     3541 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     3601 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     3661 aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc
     3721 ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc ttaaattttg
     3781 ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg gcttaactat
     3841 gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga tctcccgatc
     3901 cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3961 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4021 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4081 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4141 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4201 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4261 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4321 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4381 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4441 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4501 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4561 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4621 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4681 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4741 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4801 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4861 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4921 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4981 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5041 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5101 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5161 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5221 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5281 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5341 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5401 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5461 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5521 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5581 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5641 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5701 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5761 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5821 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5881 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5941 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6001 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6061 accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat
     6121 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     6181 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg
     6241 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     6301 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtagcacgt
     6361 gtcagtcctg ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga
     6421 actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga
     6481 agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca
     6541 cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca
     6601 cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc
     6661 ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac
     6721 tggtcaactt ggccatggtt tagttcctca ccttgtcgta ttatactat
//
