LOCUS       62056_17725        6048 bp RNA     circular SYN 30-AUG-2020
DEFINITION  Shuttle vector pNATX13-ktzT, complete sequence.
ACCESSION   MT881544
VERSION     .
KEYWORDS    .
SOURCE      
  ORGANISM  
REFERENCE   1  (bases 1 to 6048)
  AUTHORS   Chi Z, Chi Z, Kong C.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JUL-2020) Ocean University of China, Qing Dao, 
            Shandong 266000, China
REFERENCE   2  (bases 1 to 6048)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (30-JUL-2020) Ocean University of China, Qing Dao, Shandong 266000, 
            China"
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="other DNA"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /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)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3740..3757
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     protein_bind    complement(3767..3800)
                     /label=lox71
                     /note="Left element (LE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    complement(3811..3985)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(4029..4595)
                     /codon_start=1
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     protein_bind    complement(5042..5075)
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5654..5670)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgccccg gggctaaaaa ccccaacttc ggaaaggggt
     2281 gtatttatta gataaaaaac caacgccttt cggggctcct tggtgattca taataactaa
     2341 acgaatcgca tggccttgcg ccggcgatgg ttcattcaaa tttctgccct atcaactttc
     2401 gatggtagga tagtggccta ccatggtatc aacgggtaac ggggaattag ggttctattc
     2461 cggagaggga gcctgagaaa cggctaccac atccaaggaa ggcagcaggc gcgcaaatta
     2521 cccaatcccg acacggggag gtagtgacaa taaatactga tacagggctc ttttgggtct
     2581 tgtaattgga atgagtacaa tttaaatccc ttaacgagga acaattggag ggcaagtctg
     2641 gtgccagcag ccgcggtaat tccagctcca atagcgtata ttaaagttgt tgcagttaaa
     2701 aagctcgtag ttgaaccgac agttggctca tcatccgtta catcatcttt ttttctgctg
     2761 gctccgctcg gtgggctccc aacgaagcag caaaaaaagt gagagagaaa aactagcttg
     2821 gcggggcatc cgaagctaga ccctttggct cgcttagtca gtgcgccctc tcacacacac
     2881 tcaaaaaggc cacccctccc gcaccctctt ctcatcaccg tcttcatacc acggttcgtc
     2941 aagcaatcgt atctggtaag ctttgacctc ttgagcgggc tccactttgc tatttcttgg
     3001 atctgctcat tcttttcttc cttcctctct ttctaacctc tcttcagaaa gttcaaccgt
     3061 acttcactca atcttccata catcaccgtc aaacactagt atgttcgtcc ctggcccata
     3121 ccacgctcct gaggatcgct ggctcgtcga tcttgtccgt ggtcacccac tggctcagct
     3181 cgcttctaac ggtgctggtg gtgctgctcc acacatcacc cacgtcccaa tcatcgtcga
     3241 tcccgagctt gatggtcctg tcgatcgcct cgtcggcatc actctttggg gtcacatgaa
     3301 ccgtgctaac cctcactggg ctgctcttgg tggtgctgct aacgtcgtcg ctaccttcgc
     3361 cggcccaaac gcttacgtct ctcccgccgt ctacagaact gctcccgctg cccctacctg
     3421 gaacttcact tctgtccagg tccgtggcga gcttagaaag gtcgagtctg ccgacgatac
     3481 cctggctact gtccgtgcca ccgtcgctgc cctcgagtct agattcggcg ctggttggga
     3541 tatgactggc tctctggact acttccgtcg tatcctccca ggtgtcggtg ccttcagact
     3601 tcgcgtcgct gaggccgatg gcatgttcaa gctgtctcaa gagcagcaac ctgccatcag
     3661 acgccgtgtc cgccactctt tcggcggttg cgaggctacc agagctgtcg ccggccttat
     3721 ggaccgcctg cctactgagc accaccacca ccaccactaa gtcgactacc gttcgtatag
     3781 catacattat acgaagttat taattcgggg gatctggatt ttagtactgg attttggttt
     3841 taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat actaagggtt
     3901 tcttatatgc tcaacacatg agcgaaaccc tataggaacc ctaattccct tatctgggaa
     3961 ctactcacac attattatgg agaaactcga gcttgtcgat cgacagatcc cggtcggcat
     4021 ctactttatg gacatggcat ggacatgtac aaagcctgtt caccatcaga agcagtaccg
     4081 tcgtacaaag cagtatccaa accacacaag gtgaaaccca ttcttctgta agcgtggata
     4141 gctggagcgt tgacgttggt aacctccaac cacaagtgac cagcacctct ctctctagcg
     4201 aactcagtag ccaaacccat caaagctcta ccgacaccgt gacctctgtg ttctggagcg
     4261 acctcgatgt cttcaacggt caatcttctg ttccaaccgg agtaagagac aacgacgaaa
     4321 ccagccaaat caccatcgtc accgtaagcg acgaaagttc tggagtctgg atcaccgtcc
     4381 tcaccagcgt cggactcatc gtcagattca tcgtctggga aaaccttggt caatggtgga
     4441 tcgactggaa cttctctcaa agtgaaaccg tcaccagtag cggtgactct gaaaacagta
     4501 tcagtggtga aggaaccgtc caaagcctcg atagcttcag catcacctgg aacagaagtt
     4561 ctgtatctgt aagcggtatc gtccaaagtg gtcattgtga ctgaattgga tgtgttagac
     4621 aggttgatgt gaacgacagc aacagagaaa gacgcacaga gttatataca acagagcttg
     4681 atgggttgtt gtggtgtgtg aggggcttga atctttcccc aacagcttcc gacgctcggc
     4741 atgaggggca gctcaaaaaa accgcgtcac cgacgtgcta ccgagatctt cgtgctctct
     4801 cgtgtttgcc tatcaactga tatcaccaat catgagccat ctctatgtac aaactgccct
     4861 tggatcacgt gcttcactcg acatctgcac catgtatcct tcttcgaatt gctcatcttg
     4921 atcactttat tattcaagat gccttgcacc acacgacatg tctatcgatc ggcccgtaac
     4981 gccccgtgaa gttgccagaa cgcctgcctg tcccgctgtc cgatggatgc atgaaggtaa
     5041 tataacttcg tatagcatac attatacgaa cggtaggatc cccaacaggg attgccctag
     5101 taacggcgag tgaagcggca acagctcaaa tttgaaagct ggccttcggg tccgcattgt
     5161 aatttgtaga ggatgctttg ggtgaaacgc cagtctaagt tccttggaac aggacgtcat
     5221 agagggtgag aatcccgtat gtgactggaa atgttaacct atgtaaagct ccttcgacga
     5281 gtcgagttgt ttgggaatgc agctctaaat gggaggtaaa tttcttctaa agctaaatat
     5341 tggcgagaga ccgatagcgc acaagtagag tgatcgaaag atgaaaagca ctttggtaag
     5401 agagttaaaa agcacgtgaa attgttgaaa gggaagcgct tgcaatcaga cttgtttaaa
     5461 ctgttcggcc ggtcttctga ccggtttact cagtttggac aggccagcat cagtttcggc
     5521 ggccggataa aggctttggg aatgtggcct tcacttcggt gaaggtgttg tagcccaggg
     5581 tgtaatacgg ccagccggga ctgaggtccg cgcttcggct aggatgctgg cgtaatggtt
     5641 ggaattcccc gggactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     5701 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     5761 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     5821 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     5881 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     5941 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6001 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcga
//
