LOCUS       sequence-327805-        7403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human ADARB2 mammalian expression vector with 3 C-terminal FLAG 
            tags..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  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 7403)
  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-hADARB2-3xFLAG
FEATURES             Location/Qualifiers
     source          1..7403
                     /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"
     regulatory      592..601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2845..2916
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2969..2988)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2970..2988)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3017..3034)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3023..3230
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3237..3692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3374..3395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3586..3605
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3719..3823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3824..3892,3893..4681)
                     /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(4042..4061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        4775..4920
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             4963..6000
                     /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(5998..6017)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6042..6090
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6641..7229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7130..7149
                     /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 ggccaccatg
      601 gcctcggtcc tggggagcgg cagagggtct ggagggctga gcagtcaact caaatgcaag
      661 tccaagagga ggaggaggcg gaggtccaag cggaaagata aagtaagcat attgtcaacc
      721 ttcctcgctc ctttcaagca cctgagtcct ggcatcacaa acacggagga tgacgacacc
      781 ctcagtacca gcagcgcgga ggtgaaggag aaccgcaacg tgggcaacct ggccgcgcgg
      841 ccaccgccct ccggggaccg ggcccggggc ggcgcgcccg gcgcgaagag gaagcggccg
      901 ctggaggagg ggaatggggg ccacttgtgc aaactgcagc tggtctggaa gaagctgtcg
      961 tggtcggtgg cgcccaagaa cgcgctggtg cagctgcacg agctgaggcc gggcctgcag
     1021 taccggacag tgtcgcagac gggcccggtg catgccccgg tcttcgcggt agcggtggag
     1081 gtgaacgggc tcacgttcga gggcacaggc cccaccaaga agaaggccaa gatgcgcgcg
     1141 gcggagctgg cactcaggtc cttcgtgcag ttccccaacg cctgccaggc gcacctggcc
     1201 atgggcgggg gcccgggccc cggcacggac ttcacctccg accaggccga tttccccgac
     1261 acgctcttcc aggagttcga gcccccggcg ccgcgccccg gactcgcggg aggccgcccc
     1321 ggggacgccg cgcttctgtc cgcggcctac gggcgacggc ggctgctgtg ccgcgcgctg
     1381 gacctggtgg gcccgacccc cgccaccccc gcggccccgg gcgagcgcaa ccccgtggtg
     1441 ctgctgaacc gcctgcgcgc cgggctgcgc tacgtgtgtc tggcagaacc ggccgagcgg
     1501 cgcgcgcgga gcttcgtgat ggccgtgagc gtggacggca ggacgttcga gggctcgggg
     1561 cgcagcaaga agctggcccg gggtcaggcc gcgcaggccg cactgcagga gctgttcgac
     1621 atccagatgc ccggccacgc gcccggcagg gccaggagga cgccaatgcc gcaggaattc
     1681 gcagactcca tatcccagct ggtcacacag aagttccgcg aggtgacgac ggacctcacg
     1741 cccatgcacg cccgccataa agcgctggca ggaatcgtca tgaccaaagg cctggatgct
     1801 cggcaggcgc aggtcgtggc cctgtcctcg gggaccaagt gcatcagcgg cgagcacctc
     1861 agtgaccagg ggctggtggt gaatgactgc cacgcggagg tcgtggcccg gcgggcgttc
     1921 ctgcacttcc tctacacgca gctggagctg cacctgagca agcggcgcga ggactcagag
     1981 cgatcgatat tcgtgcggtt aaaagaaggt ggctaccggc tgcgagagaa catcctcttc
     2041 catctctacg tgagcacctc cccctgtgga gacgcaagac tccactctcc ctacgagatc
     2101 accacagacc tgcacagcag caaacacctc gtcaggaagt tccgcgggca cctgcgcacc
     2161 aagatcgagt ccggggaagg gacggtcccc gtgcgtggcc ccagcgcagt gcagacctgg
     2221 gacggcgtcc tgctggggga gcagctgatc accatgtcct gcacggacaa gatcgccagg
     2281 tggaacgtcc tggggctgca gggcgcgctc ctgtcccact tcgtggagcc cgtgtacctg
     2341 cagagcatcg tggtgggcag cctgcaccac acgggccacc tcgcacgcgt catgagccac
     2401 cgcatggagg gtgtcggcca gctgcccgcc tcctaccggc acaaccggcc tctcctcagc
     2461 ggcgtgagtg acaccgaggc gcgccagccg gggaagtcgc cccccttcag catgaactgg
     2521 gtcgtgggca gcgcggacct ggagattatc aacgccacca ctgggcggag gagctgtggg
     2581 ggcccatccc ggctctgcaa gcacgtgctg tctgcacggt gggcgcggct gtatggcagg
     2641 ctgagcacac ggacacccag ccctggagac acgccctcca tgtactgtga ggccaagctg
     2701 ggggcgcaca cctaccagtc tgtgaaacag cagctgttca aggcctttca gaaggctggc
     2761 ctgggcacct gggtgaggaa accaccggag cagcagcagt ttctactgac tctcggatcc
     2821 aaagcggccg caatcgatgt cgaggattac aaggatgacg acgataagga ctataaggac
     2881 gatgatgaca aggactacaa agatgatgac gataaatagg gcccggtacc ttaattaatt
     2941 aaggtaccag gtaagtgtac ccaattcgcc ctatagtgag tcgtattaca attcactcga
     3001 tcggctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3061 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3121 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3181 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga acgcgtaaat
     3241 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3301 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3361 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3421 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3481 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3541 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3601 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3661 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3721 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3781 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3841 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3901 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3961 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4021 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4081 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4141 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4201 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4261 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4321 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4381 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4441 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4501 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4561 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4621 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4681 gcctcaggat tagtgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc
     4741 ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt
     4801 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccga ggtccacttc
     4861 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
     4921 cagcgtcaac agcgtgccgc agatcccggg gggcaatgag atatgaaaaa gcctgaactc
     4981 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
     5041 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     5101 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     5161 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc
     5221 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     5281 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat
     5341 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     5401 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     5461 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     5521 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     5581 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     5641 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     5701 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     5761 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     5821 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     5881 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     5941 gatagtggaa accgacgccc cagcactcgt ccggatcggg agatggggga ggctaactga
     6001 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     6061 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     6121 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     6181 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     6241 caggccctgc catagccact ggccccgtgg gttagggacg gggtccccca tggggaatgg
     6301 tttatggttc gtgggggtta ttattttggg cgttgcgtgg ggtcaggtcc acgactggac
     6361 tgagcagaca gacccatggt ttttggatgg cctgggcatg gaccgcatgt actggcgcga
     6421 cacgaacacc gggcgtctgt ggctgccaaa cacccccgac ccccaaaaac caccgcgcgg
     6481 atttctggcg ccgccggacg aactaaacct gactacgaca tctctgcccc ttcttcgctg
     6541 gtacgaggag cgctttgcct caggtcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6601 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6661 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6721 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6781 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6841 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6901 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6961 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7021 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7081 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7141 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7201 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7261 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7321 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     7381 catagcccat atatggagtt ccg
//
