LOCUS       Exported                7393 bp ds-DNA     circular SYN 01-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS     pCMV_NESPylRS(WT)_hU6tRNAPyl
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7393)
  AUTHORS   Gemma
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 1, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     XbaI site introduced to excise the RS
            ORIGDB|GenBank
            Freezer Position: Plasmid box #15,26
FEATURES             Location/Qualifiers
     source          1..7393
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        150..529
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        530..733
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        964..982
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    1054..1104
                     /label=NES
     misc_feature    1102..2466
                     /label=PylRS(WT)
                     /note="/vntifkey=21"
     polyA_signal    2486..2710
                     /label=BGH pA
                     /label=BGH\pA
                     /note="/vntifkey=25"
     rep_origin      2756..3184
                     /label=f1 origin
                     /label=f1\origin
                     /note="/vntifkey=33"
     promoter        3198..3528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3379..3514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3577..4602
                     /codon_start=1
                     /label=Hygro(R)
                     /note="/invitrogen=1180000"
                     /note="/vntifkey=4"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4732..4853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4926..4942
                     /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(4950..4980)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4995..5016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5304..5892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6063..6923)
                     /codon_start=1
                     /label=Amp(R)
                     /note="/invitrogen=1020000"
                     /note="/vntifkey=4"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6924..7028)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    complement(7046..7117)
                     /label=tRNA Lys
                     /label=tRNA\Lys
                     /note="/vntifkey=21"
     misc_feature    complement(7125..7381)
                     /label=hU6-Promoter
                     /note="/vntifkey=21"
     promoter        complement(7126..7366)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
ORIGIN
        1 gccagatata cgccaatatt ggccattagc catattattc attggttata tagcataaat
       61 caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat
      121 tggctcatgt ccaatatgac cgccatgttg acattgatta ttgactagtt attaatagta
      181 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      241 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      301 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt
      361 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      421 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      481 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      541 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      601 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      661 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      721 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
      781 tgacctccat agaagacacc gggaccgatc cagcctccgc ggccgggaac ggtgcattgg
      841 aacgcggatt ccccgtgcca agagtgacgt aagtaccgcc tatagagtct ataggcccac
      901 ccccttggct tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
      961 acgatttagg tgacactata gaataacatc cactttgcct ttctctccac aggtgtccac
     1021 tcccaggtcc aactgcacgg aagcttcacc atggcgtgcc cggtgccgct gcagctgccg
     1081 ccgctggaac gcctgaccct ggatgataaa aaaccgctga ataccctgat ctctgctact
     1141 ggtctgtgga tgagtcgtac cggaaccatt cataaaatca aacaccacga ggttagccgt
     1201 tcgaaaatct atattgagat ggcgtgtggc gatcatctgg ttgtgaacaa tagccgctct
     1261 tctcgtacag cacgtgcact gcgtcaccac aaatatcgta aaacctgtaa acgttgccgt
     1321 gtgtccgatg aggatctgaa caaattcctg acaaaagcca atgaggacca aacaagcgtg
     1381 aaagtgaaag tcgttagcgc tcctacccgt actaaaaaag caatgccgaa atccgttgct
     1441 cgtgccccta aaccactgga aaacactgaa gcagcacagg cacagccgtc tggaagcaaa
     1501 ttctctccgg ccattcctgt ttctacccag gagtccgttt ctgttccagc aagtgtgagc
     1561 accagcatta gcagtattag caccggtgcc accgctagcg ccctggttaa aggcaatacc
     1621 aatccgatta caagcatgtc tgccccggtt caagcatcag ctccagcact gacaaaatcc
     1681 caaaccgatc gtctggaggt tctgctgaat ccgaaagacg aaatcagcct gaattccggc
     1741 aaaccgtttc gtgaactgga gagcgaactg ctgtcacgtc gtaaaaaaga cctgcaacaa
     1801 atctatgccg aagaacgtga gaactatctg gggaaactgg aacgtgaaat cacccgcttt
     1861 ttcgtggatc gtggctttct ggagatcaaa tccccgattc tgattcctct ggagtatatc
     1921 gagcgtatgg gcatcgacaa tgataccgaa ctgagcaaac aaattttccg tgtggataaa
     1981 aacttctgtc tgcgccctat gctggcacca aatctgtata actatctgcg caaactggac
     2041 cgtgccctgc ctgatcctat caaaatcttc gagatcggcc cgtgttatcg taaagagtcc
     2101 gacggtaaag aacatctgga ggagtttacc atgctgaact tttgccaaat gggttcaggt
     2161 tgtactcgtg agaacctgga aagcatcatc accgattttc tgaaccacct gggcattgac
     2221 ttcaaaattg tgggcgacag ctgtatggtg tatggcgaca ccctggatgt catgcacggc
     2281 gacctggaac tgtctagtgc cgttgtgggc ccaatcccgc tggatcgtga gtggggtatc
     2341 gacaaacctt ggatcggtgc gggttttggt ctggagcgtc tgctgaaagt aaaacacgac
     2401 ttcaagaaca tcaaacgtgc tgcacgttcc gagtcctatt acaatggtat ttctactaac
     2461 ctgtaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2521 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2581 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2641 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2701 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2821 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2881 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     2941 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3061 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3121 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3181 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca
     3241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3361 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3421 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3481 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3541 gcttgtatat ccattttcgg atctgatcag cacgtgatga aaaagcctga actcaccgcg
     3601 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     3661 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
     3721 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
     3781 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc
     3841 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
     3901 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
     3961 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     4021 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     4081 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     4141 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     4201 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     4261 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     4321 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     4381 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     4441 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     4501 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     4561 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtgct acgagatttc
     4621 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4681 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4741 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4801 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4861 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4921 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4981 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5041 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5101 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5161 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5401 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5461 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     5521 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5581 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5641 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5701 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5761 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5821 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6061 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6121 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6181 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6241 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6301 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6361 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6421 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6481 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6541 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6601 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6661 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6721 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6781 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6841 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6901 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6961 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7021 gttccgcgca catttccccg aaaaatggcg gaaaccccgg gaatctaacc cggctgaacg
     7081 gatttagagt ccattcgatc tacatgatca ggtttccggt gtttcgtcct ttccacaaga
     7141 tatataaagc caagaaatcg aaatactttc aagttacggt aagcatatga tagtccattt
     7201 taaaacataa ttttaaaact gcaaactacc caagaaatta ttactttcta cgtcacgtat
     7261 tttgtactaa tatctttgtg tttacagtca aattaattct aattatctct ctaacagcct
     7321 tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctcttcc tgcccgacct
     7381 tcgcgatgta cgg
//
