LOCUS       Exported                5387 bp ds-DNA     circular SYN 17-NOV-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5387)
  AUTHORS   Mayo Clinic Research
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 24, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5387
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(1)
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     misc_feature    60..112
                     /label=bom
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=37.5886524822695"
                     /note="/other=misc feature"
     enhancer        267..646
                     /label=CMV enhancer
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=enhancer"
     promoter        647..850
                     /label=CMV promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     LTR             846..879
                     /label=5' LTR (truncated)
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=True"
                     /note="/identity=97.059"
                     /note="/match_length=18.88888888888889"
                     /note="/other=LTR"
     promoter        895..913
                     /label=T7 promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             936..1013
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8A MTS
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1026..1091
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1095..1100
                     /codon_start=1
                     /label=2 aa linker
                     /translation="GS"
     CDS             1101..1190
                     /codon_start=1
                     /label=DddAtox 1397C
                     /translation="AIPVKRGATGETKVFTGNSNSPKSPTKGGC"
     CDS             1191..1202
                     /codon_start=1
                     /label=4 aa linker
                     /translation="SGGS"
     CDS             1203..1454
                     /codon_start=1
                     /label=UGI
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     3'UTR           1458..1605
                     /label=ATP5B 3'UTR
     promoter        1609..1630
                     /label=SV40 promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=6.197183098591549"
                     /note="/other=promoter"
     rep_origin      1651..2079
                     /direction=RIGHT
                     /label=f1 ori
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     promoter        2093..2422
                     /label=SV40 promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     promoter        2470..2517
                     /label=EM7 promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             2536..2934
                     /codon_start=1
                     /label=BSD
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             2935..2962
                     /codon_start=1
                     /label=BleoR
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=7.588075880758807"
                     /note="/other=CDS"
                     /translation="ALRGRGAGL"
     polyA_signal    3092..3222
                     /label=SV40 poly(A) signal
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=98.496"
                     /note="/match_length=97.08029197080292"
                     /note="/other=polyA signal"
     protein_bind    3284..3306
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=95.83333333333334"
                     /note="/other=protein bind"
     promoter        3310..3340
                     /label=lac promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=96.875"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     protein_bind    3355..3376
                     /label=CAP binding site
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein bind"
     CDS             complement(3389..3482)
                     /codon_start=1
                     /label=lacI
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=8.679593721144968"
                     /note="/other=CDS"
                     /translation="SAQYANRLSPRVGRFINAAGTTGFPTGKRAV"
     rep_origin      complement(3664..4252)
                     /direction=LEFT
                     /label=ori
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     misc_feature    3949
                     /label=C>T
     CDS             complement(4423..5283)
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5284..5387)
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
ORIGIN
        1 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
       61 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
      121 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
      181 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
      241 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
      301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      601 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      661 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      781 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      841 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      901 gactcactat agggagaccc aagctggcta gcaccatggc ctccgttctg accccgctgc
      961 tgctgcgtgg cctgaccggc tccgctcgtc gtctgccagt tccgcgtgcg aaaatccatt
     1021 ccctggacta caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg
     1081 acgatgacaa gatgggatcc gccattccag tgaagcgcgg cgctaccggt gaaaccaaag
     1141 tgtttaccgg taacagcaac agcccgaaga gcccgaccaa aggcggttgc tctggcggct
     1201 caactaatct gagcgacatc attgagaagg agactgggaa acagctggtc attcaggagt
     1261 ccatcctgat gctgcctgag gaggtggagg aagtgatcgg caacaagcca gagtctgaca
     1321 tcctggtgca caccgcctac gacgagtcca cagatgagaa tgtgatgctg ctgacctctg
     1381 acgcccccga gtataagcct tgggccctgg tcatccagga ttctaacggc gagaataaga
     1441 tcaagatgct gtgataaggg gtctttgtcc tctgtactgt ctctctcctt gcccctaacc
     1501 caaaaagctt catttttctg tgtaggctgc acaagagcct tgattgaaga tatattcttt
     1561 ctgaacagta tttaaggttt ccaataaaat gtacacccct cagaacttct gaggcggaaa
     1621 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     1681 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1741 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1801 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1861 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1921 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1981 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2041 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2101 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2161 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2221 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2281 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2341 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     2401 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     2461 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     2521 gtgaggaact aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga
     2581 gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct
     2641 ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct
     2701 tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt
     2761 atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag
     2821 gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg
     2881 acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcactt
     2941 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
     3001 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3061 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     3241 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgtg ccttatccgg
     3961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4261 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4321 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4381 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4441 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4501 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4561 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4621 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4681 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4741 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4801 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4861 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4921 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4981 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5041 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5101 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5161 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5221 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5281 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5341 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgc
//
