LOCUS       pMAT36                  7571 bp DNA     circular SYN 06-OCT-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7571)
  AUTHORS   Melene
  TITLE     Direct Submission
  JOURNAL   Exported Oct 6, 2022 from SnapGene 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7571
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    693..717
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    786..818
                     /label=MCS
                     /note="pUC18 multiple cloning site"
     promoter        819..837
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    838..862
                     /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)."
     RBS             877..899
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             905..1132
                     /codon_start=1
                     /label=mirA
                     /translation="MRAAINSPSLSIDTMDYQAECQFALEPSIQGLIEKAEHAGWNRQQ
                     AALAIVALASEHLTDLLTAGGLPAPDQRPLS"
     CDS             1138..1153
                     /codon_start=1
                     /label=linker
                     /note="linker"
                     /translation="GSGWS"
     CDS             1154..1225
                     /codon_start=1
                     /label=3xFLAG
                     /note="3xFLAG"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK*"
     misc_feature    1226
                     /label=insert
                     /label=nonstandard type: insert
                     /note="/source=pal"
                     /note="/type=Custom cloned insert"
     terminator      1230..1655
                     /note="terminator"
     terminator      1756..1803
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             3052..3681
                     /codon_start=1
                     /product="stability protein from Pseudomonas plasmid pVS1"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4110..5183
                     /codon_start=1
                     /product="replication protein from Pseudomonas plasmid
                     pVS1"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      5249..5443
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    5787..5927
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6113..6701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6945..7571,1..165))
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase"
                     /label=Sp/SmR
                     /note="confers resistance to spectinomycin and
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga
       61 gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac
      121 ctctgatagt tgagtcgata cttcggcgat caccgcttcc cccatgatgt ttaactttgt
      181 tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc
      241 cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca
      301 tgtcataaca agaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
      361 aggttctgga ccagttgcgt gacggcagtt acgctacttg cattacagct tacgaaccga
      421 acgaggctta tgtccactgg gttcgtgccc gaattgatca caggcagcaa cgctctgtca
      481 tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta
      541 gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct
      601 ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg
      661 agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa
      721 ttgacgctta gacaacttaa taacacattg cggacgtttt taatgtactg aattaacgcc
      781 gaattgaatt cgagctcggt acccggggat cctctagata atacgactca ctatagggga
      841 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
      901 atacatgcgg gccgcaatta attccccgtc gctttcgatc gacaccatgg actatcaggc
      961 cgagtgccaa ttcgcactgg agccttccat tcaagggctg atcgaaaaag cggaacatgc
     1021 cggctggaac cgccagcagg ccgcccttgc catcgtcgcg ctcgccagcg agcatctcac
     1081 cgatctcctg accgccggcg gcctccccgc gccggaccaa cgccccctct ccctcgaggg
     1141 tccggctggt ctggactaca aagaccatga cggtgattat aaagatcatg atatcgacta
     1201 caaagatgac gacgataaat agtaagcttg gctgttttgg cggatgagag aagattttca
     1261 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     1321 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     1381 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     1441 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1501 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     1561 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     1621 ctgacggatg gcctttttgc gtttctacaa actcttgcgg ccgcactcga gcaccaccac
     1681 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1741 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1801 ttgtaataac cgggcaggcc atgtctgccc gtatttcgcg taaggaaatc cattatgtac
     1861 tatttagtac tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag
     1921 tgcagccgtc ttctgaaaac gacatgtcgc acaagtccta agttacgcga caggctgccg
     1981 ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt
     2041 gcgactagaa ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct
     2101 atgcccgcgt cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg
     2161 ccggctgcac caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc
     2221 tggccaggat gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc
     2281 gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg
     2341 cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg
     2401 tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc
     2461 ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca
     2521 ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag
     2581 aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg
     2641 aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg
     2701 aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca
     2761 ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc
     2821 ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct
     2881 ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac
     2941 cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt
     3001 cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt
     3061 atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc
     3121 cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt
     3181 gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc
     3241 ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga
     3301 gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt
     3361 ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag
     3421 cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc
     3481 aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt
     3541 cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc
     3601 gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa
     3661 attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca
     3721 cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg
     3781 cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc
     3841 tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg
     3901 cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa
     3961 ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt
     4021 ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc
     4081 actggaaccc ccaagcccga ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac
     4141 aaatcggcgc ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc
     4201 agcggcaacg catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc
     4261 gaatccgcaa agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc
     4321 ccaagggcga cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg
     4381 atagtcgcag catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg
     4441 gcgaggtgat ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg
     4501 gcatggccag tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat
     4561 ccatgaaccg ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg
     4621 ttgcggacgt actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg
     4681 tagaaacctg cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca
     4741 agaacggccg cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg
     4801 taaagagcga aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc
     4861 gcgagatcac agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga
     4921 tcgatcccgg catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag
     4981 aagccagatg gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga
     5041 agttctgttt caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga
     5101 aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg
     5161 gcgaagcatc cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag
     5221 gggaaaaagg tcgaaaaggt ctctttcctg tggatagcac gtacattggg aacccaaagc
     5281 cgtacattgg gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt
     5341 cacacatgta agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt
     5401 taaaacttat taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca
     5461 cagccgaaga gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg
     5521 cttcgcgtcg gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca
     5581 atctaccagg gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag
     5641 gcaccctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5701 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5761 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     5821 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5881 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     5941 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6001 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6061 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6121 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6181 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6241 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6301 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6361 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6421 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6481 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6541 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6601 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6661 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6721 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatg
     6781 atatatctcc caatttgtgt agggcttatt atgcacgctt aaaaataata aaagcagact
     6841 tgacctgata gtttggctgt gagcaattat gtgcttagtg catctaatcg cttgagttaa
     6901 cgccggcgaa gcggcgtcgg cttgaacgaa tttctagcta gacattattt gccgactacc
     6961 ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc
     7021 aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac
     7081 tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg
     7141 gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc
     7201 cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt
     7261 tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct
     7321 cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct
     7381 gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc
     7441 cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc
     7501 tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca
     7561 tcaagcctta c
//
