LOCUS       sequence-175732-        5183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of p14-hARF with N-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5183)
  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 5183)
  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"; doi: 
            "10.1074/jbc.M115.663641"; journalName: "J Biol Chem"; date: 
            "2015-10-9- 9"; volume: "290"; issue: "41"; pages: "25174-87"
COMMENT     Sequence Label: FLAG-ARF
FEATURES             Location/Qualifiers
     source          1..5183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             618..641
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1207..1829
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1406..1425)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1853..1873)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1858..2187
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2038..2173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2225..2244)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2226..2244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2258..2274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2258..2274)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2271..2293)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2282..2298
                     /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)."
     promoter        complement(join(2306..2312,2313..2330,2331..2336))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2351..2372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2489..2506)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2660..3248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2740..2759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3419..4210,4211..4279))
                     /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     4042..4061
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4280..4384)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4411..4866
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4498..4517)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4708..4729
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4992..5014
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5006..5023
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5007..5023
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc taccatggac tacaaagacg atgacgacaa gcttgcggcc gacgaattcg
      661 tgcgcaggtt cttggtgacc ctccggattc ggcgcgcgtg cggcccgccg cgagtgaggg
      721 ttttcgtggt tcacatcccg cggctcacgg gggagtgggc agcgccaggg gcgcccgccg
      781 ctgtggccct cgtgctgatg ctactgagga gccagcgtct agggcagcag ccgcttccta
      841 gaagaccagg tcatgatgat gggcagcgcc cgagtggcgg agctgctgct gctccacggc
      901 gcggagccca actgcgccga ccccgccact ctcacccgac ccgtgcacga cgctgcccgg
      961 gagggcttcc tggacacgct ggtggtgctg caccgggccg gggcgcggct ggacgtgcgc
     1021 gatgcctggg gccgtctgcc cgtggacctg gctgaggagc tgggccatcg cgatgtcgca
     1081 cggtacctgc gcgcggctgc ggggggcacc agaggcagta accatgcccg catagatgcc
     1141 gcggaaggtc cctcagacat ccccgattga aagaaccaga gaggctctga gaaacctcgg
     1201 gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     1261 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     1321 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     1381 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     1441 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     1501 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     1561 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     1621 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     1681 cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca taggctacct
     1741 ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt
     1801 gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg
     1861 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     1921 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     1981 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2041 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2101 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2161 ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt
     2221 aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa
     2281 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2341 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2401 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2461 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2521 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2581 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2641 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2701 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2761 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2821 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2881 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2941 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3001 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3061 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3121 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3241 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3301 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3361 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3421 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3481 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3541 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3601 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3661 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3721 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3781 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3841 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3901 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3961 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4021 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4081 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4141 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4201 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4261 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4321 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4381 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     4441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4621 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4741 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4801 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4861 caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     4921 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     4981 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac
     5041 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
     5101 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
     5161 atgtccaata tgaccgccat gtt
//
