LOCUS       sequence-327031-        7862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human SAPS1 mammalian expression vector with C-terminal HA tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7862)
  AUTHORS   Li S, Wang L, Fu B, Berman MA, Diallo A, Dorf ME
  TITLE     TRIM65 regulates microRNA activity by ubiquitination of TNRC6.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 May 13;111(19):6970-5. doi: 
            10.1073/pnas.1322545111. Epub 2014 Apr 28.
  PUBMED    24778252
REFERENCE   2  (bases 1 to 7862)
  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: 
            "10.1073/pnas.1322545111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-05-13- 13"; volume: "111"; issue: "19"; pages: "6970-5"
COMMENT     Sequence Label: pCMV3Tag8li-hSAPS1-HA
FEATURES             Location/Qualifiers
     source          1..7862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        554..572
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3253..3279
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3304..3375
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(3428..3447)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3429..3447)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3476..3493)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3482..3689
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3696..4151)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3833..3854)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4045..4064
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4178..4282
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4283..4351,4352..5140)
                     /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(4501..4520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        5234..5379
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             5422..6459
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(6457..6476)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6501..6549
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7100..7688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7589..7608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggctcga ggccatgttt
      601 tggaagtttg acctgcacac aagctcgcac ctggacacgc tgctggagcg ggaggacctg
      661 agcctgcccg agctgctgga cgaggaagac gtgctgcagg agtgcaaggt cgtcaaccgc
      721 aagctgctgg acttcctgct gcagccaccc cacctgcaag caatggtggc ctgggtcacc
      781 caggagccgc cagatagcgg tgaggagcgg ctgcgctaca agtaccccag tgtggcctgc
      841 gagattctga cctcagatgt gccccagatc aatgatgccc tgggtgctga tgagtccctt
      901 ctgaaccggc tctacggctt cctgcagagc accggcagcc tcaacccact gctggccagc
      961 ttcttcagca aggtcatggg catcctcatc aaccgcaaga cagaccagct cgtgtccttt
     1021 cttcggaaga aggatgactt cgtggacctg ctgctgcagc acattggcac ctcggccatc
     1081 atggacctcc tgctgcgcct gctcacctgt gtggagcggc ctcagctgag gcaggatgtt
     1141 gtcaattggc tcaacgagga gaagatcgtc cagcggctga ttgagcagat ccacccgtcg
     1201 aaggatgaga atcaacattc caacgcatcc cagtccctgt gtgacatcat ccgcctgagc
     1261 cgggagcaga tgatccaagt ccaggacagc ccagagcctg accaactgct ggccaccctg
     1321 gagaagcagg agacgattga gcagctctta agcaacatgt tcgaggggga gcagagccag
     1381 tctgtcatcg tcagtgggat ccaggtgctg ctgaccctgc tggagcccag gaggccgagg
     1441 tccgagtccg tgaccgtgaa cagcttcttc agcagtgtgg atgggcagct ggagctcctg
     1501 gcccaggggg ccctggaaag cactgtgtcc agtgtgggcg ccttgcacgc cctacgcccg
     1561 cggctcagct gcttccacca gctcctgctg gagcctccca agctggagcc gctacagatg
     1621 acatggggca tgctggctcc gcctctgggc aacacgcggc tgcacgtggt caagctcctg
     1681 gccagtgccc tgagcgccaa tgatgcagcc ctgacgcacg agctcctggc actggacgtg
     1741 cccaacacca tgctggacct cttcttccat tatgtcttca acaacttctt gcatgcccaa
     1801 gtagagggat gcgtgagcac catgctgagc ttggggccac ctcctgacag cagccctgag
     1861 acgcccatcc aaaaccctgt tgtgaaacat ctgctgcagc agtgccgcct ggtggagcgg
     1921 atcctgacgt cctgggagga gaacgaccgt gtacagtgtg cgggaggccc tcggaaaggc
     1981 tacatgggtc acctgacaag agtggccggt gccctggtgc agaacacgga gaaggggccc
     2041 aatgcagagc agctgcggca gctgctgaag gagctgccca gcgagcagca ggagcagtgg
     2101 gaagccttcg tatcggggcc cctggcggag accaacaaga agaacatggt ggacctggtg
     2161 aacacccacc acctacactc ctccagtgac gatgaggacg accggctcaa ggagttcaac
     2221 ttccctgagg aggctgtgct gcagcaggcc ttcatggact tccagatgca gcgcatgacc
     2281 tctgccttca ttgaccactt cggcttcaat gatgaggagt ttggggagca ggaggagagt
     2341 gtgaacgcac cttttgacaa gacagccaac atcaccttct ccctcaatgc tgacgatgag
     2401 aaccccaacg ccaacctact tgagatatgc tacaaggacc gcatccagca gtttgatgat
     2461 gatgaggaag aggaggacga ggaagaggcc cagggctcag gggagtctga tggagaagat
     2521 ggcgcctggc agggcagcca gctggccagg ggggcccgtc tgggccagcc acctggtgtc
     2581 cggagtggag gcagcacaga cagtgaggac gaagaagagg aggacgagga ggaggaggaa
     2641 gacgaggagg gcattggctg tgcagcccgt ggaggggcca cccctctgtc ctaccccagc
     2701 cctggccctc agcctccagg ccccagctgg acagccacct ttgacccagt gcctacagat
     2761 gccccgacca gcccccgagt ctccggggag gaagagctgc acactgggcc tccagcccca
     2821 caggggcccc tcagtgtgcc ccagggcctc cccactcaga gcctggccag ccctcctgcc
     2881 cgtgacgccc tgcagctcag gtctcaggac cccacacccc cctcagcacc tcaggaagcc
     2941 acagaaggca gcaaagtcac ggagccctca gccccttgcc aggccttggt tagcatcggg
     3001 gaccttcagg ccaccttcca cgggatccgt tctgccccca gctcctcgga cagtgcaacc
     3061 agagacccct ctacctctgt cccagcctcc ggggcccacc agccccccca gaccacagaa
     3121 ggggagaaga gcccagagcc cttggggctt ccccaaagcc agagtgccca ggccctcacg
     3181 cctcctccga tacccaatgg ctctgccccg gaagggcctg catccccagg ctcccaagcg
     3241 gccgcaatcg attacccata cgatgttcca gattacgctc tttaggaatt cttcgatgtc
     3301 gaggattaca aggatgacga cgataaggac tataaggacg atgatgacaa ggactacaaa
     3361 gatgatgacg ataaataggg cccggtacct taattaatta aggtaccagg taagtgtacc
     3421 caattcgccc tatagtgagt cgtattacaa ttcactcgat cggctcgctg atcagcctcg
     3481 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3541 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3601 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3661 tgggaagaca atagcaggca tgctggggaa cgcgtaaatt gtaagcgtta atattttgtt
     3721 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3781 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3841 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3901 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3961 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4021 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4081 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4141 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4201 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4261 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4321 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4381 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4441 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4501 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4561 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4621 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4681 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4741 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4801 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4861 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4921 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4981 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5041 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5101 acagatcgct gagataggtg cctcactgat taagcattgg cctcaggatt agtgcttcat
     5161 ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt
     5221 ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac
     5281 gcagatgcag tcggggcggc gcggtccgag gtccacttcg catattaagg tgacgcgtgt
     5341 ggcctcgaac accgagcgac cctgcagcga cccgcttaac agcgtcaaca gcgtgccgca
     5401 gatcccgggg ggcaatgaga tatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     5461 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     5521 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     5581 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     5641 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     5701 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     5761 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     5821 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     5881 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     5941 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     6001 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     6061 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     6121 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     6181 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     6241 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     6301 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     6361 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     6421 agcactcgtc cggatcggga gatgggggag gctaactgaa acacggaagg agacaatacc
     6481 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     6541 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     6601 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     6661 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagccactg
     6721 gccccgtggg ttagggacgg ggtcccccat ggggaatggt ttatggttcg tgggggttat
     6781 tattttgggc gttgcgtggg gtcaggtcca cgactggact gagcagacag acccatggtt
     6841 tttggatggc ctgggcatgg accgcatgta ctggcgcgac acgaacaccg ggcgtctgtg
     6901 gctgccaaac acccccgacc cccaaaaacc accgcgcgga tttctggcgc cgccggacga
     6961 actaaacctg actacgacat ctctgcccct tcttcgctgg tacgaggagc gctttgcctc
     7021 aggtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7081 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7141 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7201 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7261 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7321 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7381 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7441 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7501 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7561 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7621 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7681 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7741 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     7801 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     7861 cg
//
