LOCUS       sequence-206330-        6846 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses epitope-tagged wild-type human TTP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6846)
  AUTHORS   Coelho MA, de Carne Trecesson S, Rana S, Zecchin D, Moore C, 
            Molina-Arcas M, East P, Spencer-Dene B, Nye E, Barnouin K, Snijders 
            AP, Lai WS, Blackshear PJ, Downward J
  TITLE     Oncogenic RAS Signaling Promotes Tumor Immunoresistance by 
            Stabilizing PD-L1 mRNA.
  JOURNAL   Immunity. 2017 Dec 19;47(6):1083-1099.e6. doi: 
            10.1016/j.immuni.2017.11.016. Epub 2017 Dec 12.
  PUBMED    29246442
REFERENCE   2  (bases 1 to 6846)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.immuni.2017.11.016"; journalName: "Immunity"; date: 
            "2017-12-19- 19"; volume: "47"; issue: "6"; pages: "1083-1099.e6"
COMMENT     Sequence Label: pcDNA3 Myc2- Hs TTP WT
FEATURES             Location/Qualifiers
     source          1..6846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     702..721
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        702..720
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             768..797
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             798..827
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2693..2710)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2699..2923
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2969..3397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3056..3075)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3266..3287
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3406..3426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3411..3740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3591..3726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3653..3672
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3788..3835
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3854..4228
                     /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"
     polyA_signal    4358..4479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4395..4414)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4449..4468
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4528..4544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4528..4544)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4541..4563)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4552..4568
                     /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(4576..4582,4583..4600,4601..4606))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4621..4642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4759..4776)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4930..5518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5010..5029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5689..6480,6481..6549))
                     /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     6312..6331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6550..6654)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6729..6748)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
       61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      661 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
      721 gagacccaag ctggctagcg tttaaactta agcttggtac caccatggag cagaagctga
      781 tctcagagga ggatctggag cagaagctga tctcagagga ggatctggat atccagcaca
      841 gtggcggccg cgccaaccgt tacaccatgg atctgactgc catctacgag gtgagtcccc
      901 gccgcacggc atccccggta cctgcatgcc tgagtccgag tccccacctc tctagcgccg
      961 caaactccag cccgggacgc ttgcctccct tctccaactg gggctcccta gcgccgcgcc
     1021 ctccagcctg gggcccctgc ctcccgctca gaccagcttg gtgatttgga ggtgaaaatg
     1081 gaacccgcga cacccggctc ttcgctcaaa catgggtggg gcggcccatg caagtggaaa
     1141 gtcggagaac ttttctcaga ccgaggctgc ctggaggcgg aagtggcccc catacctggc
     1201 tcacccctag tcgttgctga gggcgtggtt ttgcgcggag gcgtctctgg ggctgaagtc
     1261 tcagggtggg gggatccgac ttctgtctct ccagtccctg accgtagaga cagagaaccc
     1321 taaaaccgaa gcaatccgga cttccaggtc aactttgccc ggtttctcca gttgtgaaac
     1381 tggagatccc gacgcgtggg tcatatccgg ggaggacaag agacccaaaa ttgggaaaca
     1441 gtggtgcgcc ctgacttcgg ggtccccctc ttggtccagc cggggaagcc gggattcctg
     1501 ggtccctcgg gataaggcct cggtggtggg taaactcaga acctccaact ctgggttcct
     1561 ggcatccgga acccaggggt ttctgcgggc gggtggggct caggcgggga gcccacaaac
     1621 cggcctggca agctctagtt ccctgcagct ggggtggggc gtcgccctgc attttcaggt
     1681 gccttaaccg acccatttcc gcagagcctc ctgtcgctga gccctgacgt gcccgtgcca
     1741 tccgaccatg gagggactga gtccagccca ggctggggct cctcgggacc ctggagcctg
     1801 agcccctccg actccagccc gtctggggtc acctcccgcc tgcctggccg ctccaccagc
     1861 ctagtggagg gccgcagctg tggctgggtg cccccacccc ctggcttcgc accgctggct
     1921 ccccgcctgg gccctgagct gtcaccctca cccacttcgc ccactgcaac ctccaccacc
     1981 ccctcgcgct acaagactga gctatgtcgg accttctcag agagtgggcg ctgccgctac
     2041 ggggccaagt gccagtttgc ccatggcctg ggcgagctgc gccaggccaa tcgccacccc
     2101 aaatacaaga cggaactctg tcacaagttc tacctccagg gccgctgccc ctacggctct
     2161 cgctgccact tcatccacaa ccctagcgaa gacctggcgg ccccgggcca ccctcctgtg
     2221 cttcgccaga gcatcagctt ctccggcctg ccctctggcc gccggacctc accaccacca
     2281 ccaggcctgg ccggcccttc cctgtcctcc agctccttct cgccctccag ctccccacca
     2341 ccacctgggg accttccact gtcaccctct gccttctctg ctgcccctgg cacccccctg
     2401 gctcgaagag accccacccc agtctgttgc ccctcctgcc gaagggccac tcctatcagc
     2461 gtctgggggc ccttgggtgg cctggttcgg accccctctg tacagtccct gggatccgac
     2521 cctgatgaat atgccagcag cggcagcagc ctggggggct ctgactctcc cgtcttcgag
     2581 gcgggagttt ttgcaccacc ccagcccgtg gcagcccccc ggcgactccc catcttcaat
     2641 cgcatctctg tttctgagtg atctagaggg cccgtttaaa cccgctgatc agcctcgact
     2701 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2761 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2821 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2881 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2941 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3001 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3061 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3121 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3181 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3241 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3301 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3361 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3421 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3481 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3601 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3661 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3721 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3781 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3841 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     3901 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     3961 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     4021 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     4081 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     4141 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     4201 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     4261 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4321 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4381 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4441 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4501 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4561 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4621 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4681 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4741 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4801 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4861 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5341 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5641 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5701 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5761 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5821 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5881 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5941 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6001 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6061 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6121 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6181 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6241 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6301 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6361 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6421 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6481 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6541 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6601 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6661 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     6721 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     6781 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     6841 accgac
//
