LOCUS       Exported                2961 bp ds-DNA     circular SYN 29-OCT-2024
DEFINITION  [pL0-G] Gal4-NLS-HA-VP64.
ACCESSION   urn.local.jones.umm-8qj5t5f
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2961)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 31, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Imported using the Genbank importer. File name: 2 documents from 
            Activators.gb
FEATURES             Location/Qualifiers
     source          1..2961
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(17..38)
                     /label=pHAGE-A7050-6550
                     /note="Geneious type: Promoter_prokaryotic"
                     /note="/ApEinfo_fwdcolor=#b4abac"
                     /note="/ApEinfo_revcolor=#b4abac"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pTK143 1732\-\pAAVio-CA; 
                     pTW073(pTK143 no IRES GFP)"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/vntifkey=28"
     primer_bind     222..241
                     /label=RepO-R
                     /note="/ApEinfo_fwdcolor=#ff9ccd"
                     /note="/ApEinfo_revcolor=#ff9ccd"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pTW037 DI pDEST"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    261..285
                     /label=pHAGE-S7501-8000
                     /note="Geneious type: Promoter_prokaryotic"
                     /note="/ApEinfo_fwdcolor=#b4abac"
                     /note="/ApEinfo_revcolor=#b4abac"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pTK143 1732\-\pAAVio-CA; 
                     pTW073(pTK143 no IRES GFP)"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/vntifkey=28"
     misc_feature    complement(528..549)
                     /label=pHAGE-A7550-7050
                     /note="Geneious type: Promoter_prokaryotic"
                     /note="/ApEinfo_fwdcolor=#ff9ccd"
                     /note="/ApEinfo_revcolor=#ff9ccd"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pTK143 1732\-\pAAVio-CA; 
                     pTW073(pTK143 no IRES GFP)"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/vntifkey=28"
     primer_bind     530..550
                     /label=TW_HDV-seq-R
                     /note="/ApEinfo_fwdcolor=#84b0dc"
                     /note="/ApEinfo_revcolor=#84b0dc"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pTW031 short HDV Active (-); 
                     pTW037DI pDEST; pTW039 Genomic HDV Active (-); pTW040 
                     GenomicHDV Inactive (-)"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/created_by=User"
     misc_feature    complement(537..548)
                     /label=DrdI, AasI, DseDI
                     /note="Geneious type: restriction site"
                     /note="/ApEinfo_fwdcolor=#75c6a9"
                     /note="/ApEinfo_revcolor=#75c6a9"
                     /note="/Recognition_pattern=GACNNNN^NNGTC"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pTW031 short HDV Active (-); 
                     pTW039Genomic HDV Active (-); pTW040 Genomic HDV Inactive 
                     (-)"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/annotation_group=GACNNNNNNGTC"
     misc_feature    complement(557..568)
                     /label=BdaI
                     /note="Geneious type: restriction site"
                     /note="/ApEinfo_fwdcolor=#faac61"
                     /note="/ApEinfo_revcolor=#faac61"
                     /note="/Recognition_pattern=(10/12)TGANNNNNNTCA(12/10)"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pTW031 short HDV Active (-); 
                     pTW039Genomic HDV Active (-); pTW040 Genomic HDV Inactive 
                     (-)"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/annotation_group=TGANNNNNNTCA"
     rep_origin      589
                     /label=ORI
                     /note="/ApEinfo_fwdcolor=#b7e6d7"
                     /note="/ApEinfo_revcolor=#b7e6d7"
     primer_bind     660..680
                     /label=TW_lvl0_seq_Rev1
                     /note="/ApEinfo_fwdcolor=#ff9ccd"
                     /note="/ApEinfo_revcolor=#ff9ccd"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
     primer_bind     689..708
                     /label=G00101
     misc_feature    763..777
                     /label=15 bp SapI Overhang
                     /note="/ApEinfo_fwdcolor=#9eafd2"
                     /note="/ApEinfo_revcolor=#9eafd2"
                     /note="/Transferred_From=pJB770"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     primer_bind     824..842
                     /label=RJ_00029Fa
     protein_bind    830..835
                     /label=BsaI Bind
                     /note="/ApEinfo_fwdcolor=#d6b295"
                     /note="/ApEinfo_revcolor=#d6b295"
                     /note="/created_by=User"
     misc_feature    830..835
                     /label=BsaI
                     /note="Geneious type: restriction site"
                     /note="/ApEinfo_fwdcolor=#d6b295"
                     /note="/ApEinfo_revcolor=#d6b295"
                     /note="/Recognition_pattern=GGTCTC(1/5)"
                     /note="/annotation_group=GGTCTC"
     misc_feature    837..840
                     /label=pL0 5-G
                     /label=nonstandard type:  Gg_overhang
                     /note="/ApEinfo_fwdcolor=#75c6a9"
                     /note="/ApEinfo_revcolor=#75c6a9"
                     /note="/created_by=User"
     misc_feature    841..846
                     /label=Kozak
                     /label=nonstandard type:  motif
                     /note="/ApEinfo_fwdcolor=#ffef86"
                     /note="/ApEinfo_revcolor=#ffef86"
                     /note="/created_by=User"
     CDS             847..1548
                     /codon_start=1
                     /label=Gal4-NLS-HA-VP64
                     /note="/ApEinfo_fwdcolor=#84b0dc"
                     /note="/ApEinfo_revcolor=#84b0dc"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSMGPKKKRKVGSGYPYD
                     VPDYAGSGEASGSGRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALD
                     DFDLDMLINSR"
     misc_feature    850..1287
                     /label=Gal4 DBD
                     /label=nonstandard type:  domain
                     /note="/ApEinfo_fwdcolor=#d59687"
                     /note="/ApEinfo_revcolor=#d59687"
                     /note="/created_by=User"
     misc_feature    877..960
                     /label=Zn(2)-C6fungal-type.{ECO:0000255|PROSITE-ProRul
                     /label=Zn(2)-C6fungal-type.{ECO:0000255|PROSITE-ProRule:PRU
                     00227}.
                     /note="Geneious type: dna_binding_domain"
                     /note="/ApEinfo_fwdcolor=#c7b0e3"
                     /note="/ApEinfo_revcolor=#c7b0e3"
                     /note="/Transferred_From=Gal4 DBD (pBjh_4067)"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     protein_bind    994..1125
                     /label=Leucine Zipper
                     /note="/ApEinfo_fwdcolor=#d6b295"
                     /note="/ApEinfo_revcolor=#d6b295"
                     /note="/created_by=jones"
     misc_feature    1294..1314
                     /label=NLS
                     /label=nonstandard type:  motif
                     /note="/ApEinfo_fwdcolor=#ff9ccd"
                     /note="/ApEinfo_revcolor=#ff9ccd"
                     /note="/Transferred_From=Digestion of pRJ_00009 PCR Product
                     -Fragment 2, Digestion of pRJ_00010 PCR Product - 
                     Fragment2, Digestion of pRJ_00011 PCR Product - Fragment 
                     2,Digestion of pRJ_00012 PCR Product - Fragment 2, 
                     Digestionof pRJ_00013 PCR Product - Fragment 2, 
                     pRJ_00009,pRJ_00009 PCR Product, pRJ_00010, pRJ_00010 PCR 
                     Product,pRJ_00011, pRJ_00011 PCR Product, pRJ_00012, 
                     pRJ_00012 PCRProduct, pRJ_00013, pRJ_00013 PCR Product, 
                     pRJ_00095,pRJ_00096, pRJ_00097, pRJ_00098, pRJ_00099"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     primer_bind     1294..1314
                     /label=RJ_00068Fa
     misc_feature    1315..1323
                     /label=GSG
                     /label=nonstandard type:  motif
                     /note="/ApEinfo_fwdcolor=#d59687"
                     /note="/ApEinfo_revcolor=#d59687"
                     /note="/created_by=User"
     sig_peptide     1324..1350
                     /label=HA-tag
                     /note="/ApEinfo_fwdcolor=#faac61"
                     /note="/ApEinfo_revcolor=#faac61"
                     /note="/Transferred_From=(4) Digestion of pRJ_00095 - 
                     Fragment1, (4) Digestion of pRJ_00096 - Fragment 1, (4) 
                     Digestionof pRJ_00097 - Fragment 1, (4) Digestion of 
                     pRJ_00098 -Fragment 1, (4) Digestion of pRJ_00099 - 
                     Fragment 1,Digestion of pRJ_00095 - Fragment 1, Digestion 
                     ofpRJ_00096 - Fragment 1, Digestion of pRJ_00097 - 
                     Fragment1, Digestion of pRJ_00098 - Fragment 1, Digestion 
                     ofpRJ_00099 - Fragment 1, pRJ_00095, pRJ_00096, 
                     pRJ_00097,pRJ_00098, pRJ_00099"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    1336..1350
                     /label=Infusion
                     /label=nonstandard type:  Gibson_overhang
                     /note="/ApEinfo_fwdcolor=#f58a5e"
                     /note="/ApEinfo_revcolor=#f58a5e"
                     /note="/created_by=User"
     misc_feature    1351..1359
                     /label=GSG
                     /label=nonstandard type:  motif
                     /note="/ApEinfo_fwdcolor=#f8d3a9"
                     /note="/ApEinfo_revcolor=#f8d3a9"
                     /note="/created_by=User"
     misc_feature    1363..1377
                     /label=Linker
                     /label=nonstandard type:  motif
                     /note="/ApEinfo_fwdcolor=#c6c9d1"
                     /note="/ApEinfo_revcolor=#c6c9d1"
                     /note="/created_by=User"
     misc_feature    1378..1545
                     /label=VP64
                     /label=nonstandard type:  domain
                     /note="/ApEinfo_fwdcolor=#75c6a9"
                     /note="/ApEinfo_revcolor=#75c6a9"
                     /note="/Transferred_From=Digestion of pRJ_00010 PCR Product
                     -Fragment 2, Digestion of pRJ_00011 PCR Product - 
                     Fragment2, pRJ_00010, pRJ_00010 PCR Product, pRJ_00011, 
                     pRJ_00011PCR Product, pRJ_00096, pRJ_00097"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    1384..1416
                     /label=VP16min
                     /label=nonstandard type:  domain
                     /note="/ApEinfo_fwdcolor=#b1ff67"
                     /note="/ApEinfo_revcolor=#b1ff67"
                     /note="/Transferred_From=VP64 (pBjh_4067)"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
     misc_feature    1423..1455
                     /label=VP16min
                     /label=nonstandard type:  domain
                     /note="/ApEinfo_fwdcolor=#d6b295"
                     /note="/ApEinfo_revcolor=#d6b295"
                     /note="/Transferred_From=VP64 (pBjh_4067)"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
     misc_feature    1462..1494
                     /label=VP16min
                     /label=nonstandard type:  domain
                     /note="/ApEinfo_fwdcolor=#75c6a9"
                     /note="/ApEinfo_revcolor=#75c6a9"
                     /note="/Transferred_From=VP64 (pBjh_4067)"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
     misc_feature    1501..1533
                     /label=VP16min
                     /label=nonstandard type:  domain
                     /note="/ApEinfo_fwdcolor=#75c6a9"
                     /note="/ApEinfo_revcolor=#75c6a9"
                     /note="/Transferred_From=VP64 (pBjh_4067)"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
     primer_bind     complement(1522..1545)
                     /label=RJ_00050Rc
     misc_feature    1549..1552
                     /label=pL0 G-3
                     /label=nonstandard type:  Gg_overhang
                     /note="/ApEinfo_fwdcolor=#f8d3a9"
                     /note="/ApEinfo_revcolor=#f8d3a9"
                     /note="/created_by=User"
     misc_feature    complement(1554..1559)
                     /label=BsaI
                     /note="Geneious type: restriction site"
                     /note="/ApEinfo_fwdcolor=#85dae9"
                     /note="/ApEinfo_revcolor=#85dae9"
                     /note="/Recognition_pattern=GGTCTC(1/5)"
                     /note="/annotation_group=GGTCTC"
     protein_bind    complement(1554..1559)
                     /label=BsaI Bind
                     /note="/ApEinfo_fwdcolor=#b1ff67"
                     /note="/ApEinfo_revcolor=#b1ff67"
                     /note="/created_by=User"
     misc_feature    1709..1732
                     /label=Pamp
                     /note="Geneious type: Promoter_prokaryotic"
                     /note="/ApEinfo_fwdcolor=#c6c9d1"
                     /note="/ApEinfo_revcolor=#c6c9d1"
                     /note="/created_by=User"
     primer_bind     complement(1724..1744)
                     /label=JH856R
     misc_feature    1779..2791
                     /label=Spec
                     /note="/ApEinfo_fwdcolor=#f58a5e"
                     /note="/ApEinfo_revcolor=#f58a5e"
     primer_bind     complement(2252..2275)
                     /label=RJ_00189Ra
     primer_bind     2261..2281
                     /label=RJ_00190Fa
     rep_origin      complement(join(2886..2961,1..607))
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="/ApEinfo_fwdcolor=#84b0dc"
                     /note="/ApEinfo_revcolor=#84b0dc"
                     /note="/created_by=User"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tcactctgtg gtctcaaggt
      841 gccaccatga agctactgtc ctctatcgaa caagcatgcg atatttgccg acttaaaaag
      901 ctcaagtgct ccaaagaaaa accgaagtgc gccaagtgtc tgaagaacaa ctgggagtgt
      961 cgctactctc ccaaaaccaa aaggtccccg ctgactaggg cacatctgac agaagtggaa
     1021 tcaaggctag aaagactgga acagctattt ctactgattt ttcctcgaga ggaccttgac
     1081 atgattttga aaatggattc tttacaggat ataaaagcat tgttaacagg attatttgta
     1141 caagataatg tgaataaaga tgccgtcaca gatagattgg cttcagtgga gactgatatg
     1201 cctctaacat tgagacagca tagaataagt gcgacatcat catcggaaga gagtagtaac
     1261 aaaggtcaaa gacagttgac tgtatcgatg ggccccaaga agaagaggaa ggtgggatct
     1321 ggctacccat acgacgtgcc agattacgcc ggatcaggtg aggccagcgg ttccggacgg
     1381 gctgacgcat tggacgattt tgatctggat atgctgggaa gtgacgccct cgatgatttt
     1441 gaccttgaca tgcttggttc ggatgccctt gatgactttg acctcgacat gctcggcagt
     1501 gacgcccttg atgatttcga cctggacatg ctgattaact ctagatgagc tttgagacca
     1561 cgaagtggct cttcagtgga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     1621 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     1681 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1741 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gcgctcacgc aactggtcca
     1801 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     1861 actgtttttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     1921 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     1981 taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     2041 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     2101 cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc tggttacggt
     2161 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     2221 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     2281 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     2341 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     2401 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     2461 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     2521 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     2581 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     2641 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     2701 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc attacgtaaa
     2761 aggcgagatc accaaggtag tcggcaaata actgtcagac caagtttact catatatact
     2821 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2881 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2941 agaaaagatc aaaggatctt c
//
