LOCUS       sequence-323281-        3298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VP16 activation domain, requires assembly with TL2-TL4 linker and 
            TL1 DNA binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3298)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 3298)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pVP16 AD
FEATURES             Location/Qualifiers
     source          1..3298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..75
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        290..394
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(395..463,464..1255)
                     /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(613..632)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1426..2014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1915..1934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2168..2185
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2302..2323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2338..2343,2344..2361,2362..2368)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2376..2392
                     /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     2381..2403
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2400..2416
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2400..2416
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2434..2452
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2434..2451
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2555..2788
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDTADFEFEQMFTDALGIDEYGG"
     regulatory      2829..2838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2833..2842
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2872..2891)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2873..2891)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2898..2915)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2898..2914)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2907..2929)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(3055..3298,1..212)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3142..3161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
       61 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      121 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      181 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
      241 ctgtgcggta tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      301 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      361 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      421 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
      481 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
      541 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
      601 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
      661 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
      721 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
      781 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
      841 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
      901 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
      961 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1021 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1081 tgctgataaa tctggagccg gtgagcgtgg gtcgcgcggt atcattgcag cactggggcc
     1141 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1201 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1261 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1321 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1381 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1441 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1501 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1561 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1621 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1681 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1741 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1801 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1861 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1921 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1981 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2041 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2101 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2161 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     2221 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2281 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     2341 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     2401 aggaaacagc tatgaccatg attacgccaa gctatttagg tgacactata gaatactcaa
     2461 gctatgcatc caacgcgttg ggagctctcc catatggtcg acctgcaggc ggccgcgaat
     2521 tcactagtga ttgggcgatg agtggtctcg ctcggccccc ccgaccgatg tcagcctggg
     2581 ggacgagctc cacttagacg gcgaggacgt ggccatggcg catgccgacg cgctagacga
     2641 tttcgatctg gacatgttgg gggacgggga ttccccgggg ccgggattta ccccccacga
     2701 ctccgccccc tacggcgctc tggatacggc cgacttcgag tttgagcaga tgtttaccga
     2761 tgcccttgga attgacgagt acggtgggta agcttcgaga ccactcatcg cccaatcgaa
     2821 ttcccgcggc cgccatggcg gccgggagca tgcgacgtcg ggcccaattc gccctatagt
     2881 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     2941 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3001 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     3061 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3121 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3181 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3241 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccagc
//
