LOCUS       Exported                3807 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  mammalian expression of p14-hARF linked to VP16 activation domain.
ACCESSION   .
VERSION     .
KEYWORDS    VP-ARF
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3807)
  AUTHORS   Minges JT, Grossman G, Zhang P, Kafri T, Wilson EM
  TITLE     Post-translational Down-regulation of Melanoma Antigen-A11 
            (MAGE-A11) by Human p14-ARF Tumor Suppressor.
  JOURNAL   J Biol Chem. 2015 Oct 9;290(41):25174-87. doi: 
            10.1074/jbc.M115.663641. Epub 2015 Sep 1.
  PUBMED    26330556
REFERENCE   2  (bases 1 to 3807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3807
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     68..86
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(124..146)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     246..265
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        346..364
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     346..363
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(374..394)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     613..632
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     regulatory      702..711
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             714..734
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             735..968
                     /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
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     polyA_signal    1662..1796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1699..1718)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1753..1772
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1806..1825)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1807..1825)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1912..1929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2083..2671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2163..2182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2842..3702)
                     /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     3465..3484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3703..3807)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
      121 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
      181 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
      241 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga
      301 acgcggctac aattaataca taaccttatg tatcatacac atacgattta ggtgacacta
      361 tagaactcga tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ggaaagtccc caggctcccc
      481 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
      541 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
      601 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
      661 gtagtgaaga ggcttttttg gaggagatct aagctagcgc cgccaccatg ggccctaaaa
      721 agaagcgtaa agtcgccccc ccgaccgatg tcagcctggg ggacgagctc cacttagacg
      781 gcgaggacgt ggcgatggcg catgccgacg cgctagacga tttcgatctg gacatgttgg
      841 gggacgggga ttccccgggt ccgggattta ccccccacga ctccgccccc tacggcgctc
      901 tggatatggc cgacttcgag tttgagcaga tgtttaccga tgcccttgga attgacgagt
      961 acggtgggga attcgtgcgc aggttcttgg tgaccctccg gattcggcgc gcgtgcggcc
     1021 cgccgcgagt gagggttttc gtggttcaca tcccgcggct cacgggggag tgggcagcgc
     1081 caggggcgcc cgccgctgtg gccctcgtgc tgatgctact gaggagccag cgtctagggc
     1141 agcagccgct tcctagaaga ccaggtcatg atgatgggca gcgcccgagt ggcggagctg
     1201 ctgctgctcc acggcgcgga gcccaactgc gccgaccccg ccactctcac ccgacccgtg
     1261 cacgacgctg cccgggaggg cttcctggac acgctggtgg tgctgcaccg ggccggggcg
     1321 cggctggacg tgcgcgatgc ctggggccgt ctgcccgtgg acctggctga ggagctgggc
     1381 catcgcgatg tcgcacggta cctgcgcgcg gctgcggggg gcaccagagg cagtaaccat
     1441 gcccgcatag atgccgcgga aggtccctca gacatccccg attgaaagaa ccagagaggc
     1501 tctgagaaac ctcgggatcc gtcgacgcgt ctgcagaagc ttctagataa gtaatgatca
     1561 taatcagcca tatcacatct gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1621 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     1681 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1741 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgcc
     1801 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     1861 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3781 gaaaaataaa caaatagggg ttccgcg
//
