LOCUS       40924_2433        7272 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p17GFP(lox-Sm-E-Kn)-Tc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7272)
  AUTHORS   Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV.
  TITLE     Genetic tools for C. glutamicum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7272)
  AUTHORS   Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-DEC-2014) Lab3, AGRI, 1st Dorozhny Proezd, 1-1, Moscow
            117545, Russia
REFERENCE   3  (bases 1 to 7272)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7272)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-DEC-2014) Lab3, AGRI, 1st Dorozhny Proezd, 1-1, Moscow 117545, 
            Russia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  complement(56..173)
                     /mobile_element_type="transposon:Mu bacteriophage attR"
                     /label=ttR
     protein_bind    complement(280..313)
                     /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)."
     protein_bind    359..392
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(586..1377)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    1738..1785
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      complement(1991..2138)
                     /regulatory_class="enhancer"
     CDS             complement(2217..3053)
                     /codon_start=1
                     /gene="strB"
                     /product="StrB"
                     /label=strB
                     /note="streptomycin phosphotransferase"
                     /protein_id="AKN19595.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     gene            complement(2217..3053)
                     /gene="strB"
                     /label=strB
     CDS             complement(3053..3856)
                     /codon_start=1
                     /gene="strA"
                     /product="StrA"
                     /label=strA
                     /note="streptomycin phosphotransferase"
                     /protein_id="AKN19596.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     gene            complement(3053..3856)
                     /gene="strA"
                     /label=strA
     protein_bind    complement(3926..3959)
                     /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)."
     CDS             complement(3965..4657)
                     /label=ZsGreen
                     /note="Zoanthus green fluorescent protein (Matz et al.,
                     1999)"
     primer_bind     complement(4889..4905)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5108..5167
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     mobile_element  complement(5179..5396)
                     /mobile_element_type="transposon:Mu bacteriophage attL"
                     /label=ttL
     rep_origin      complement(5526..5918)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(6008..7210)
                     /label=TcR
                     /note="tetracycline efflux protein"
     protein_bind    complement(7219..7237)
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 atcgatagtt tccttgcatg aatccataaa aggcgcctgt agtgccattt acccctgaag
       61 cggcgcacga aaaacgcgaa agcgtttcac gataaatgcg aaaactttag ctttcgcgct
      121 tcaaatgaaa cagatgtatt aattactgct ttttattcat tacatgggga tcttgaaagc
      181 tggagagaaa aatgaataga tactaaaaag ccggagcagt ctcccgctcc ggcttcacaa
      241 ggcaatcgcc ttgcagcgaa acacaattac tcttatcgct accgttcgta tagcatacat
      301 tatacgaagt tatgattcca gtgcgatttt gatcatctgc atccttagtt cctattccga
      361 agttcctatt ctctagaaag tataggaact tcagagcgct tttgaagctg gggtgggcga
      421 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
      481 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
      541 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
      601 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
      661 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
      721 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
      781 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
      841 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
      901 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
      961 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     1021 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     1081 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     1141 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     1201 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     1261 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     1321 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     1381 aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg
     1441 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     1501 agctggcaat tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt
     1561 aagcccactg caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc
     1621 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtgtt
     1681 ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg cagcgtgagg ggatcttgaa
     1741 gttcctattc cgaagttcct attctctaga aagtatagga acttcgaagc agctccagcc
     1801 tacacaatcg ctcaagacgt gtaatgctgc aagtcgacca tcgacatcac atcgtattca
     1861 acagctttcc cccccttgac accttccttt ttttgcttcg tccagccttg aacatttgct
     1921 cgatagtgaa caccagcaac agatccaggc ataccatcag cagccataat ttcttgcggc
     1981 gaacaccaaa ttgacttttc agtattattc ttttctataa agttactttt caaaatttaa
     2041 actccttatt tatcaacgcg ttaatcagta atcaaaggaa tttaccaaaa agcagcttta
     2101 caaaaagctt ttcagtaatt atctttttag taagctagct aagtttttac acttagttaa
     2161 attgctaact ttatagatta caaaacttag gagggttttt aaatgtggat ccccatctag
     2221 tatgacgtct gtcgcacctg cttgatcgcg gccgcgatag ctagatcgcg ttgctcctct
     2281 tctccatccg cgttccaagc tgcggaaagg cacccataag cgtacgcctg gtcgagcagg
     2341 cgacgcggat cgacgtccag cgcacgagag aatgcgtccg ccatctgtgc aatgcgtcta
     2401 ggatcgagac aaaggtcgtc tctgtcagcc ggatcgtaga acatattggc ggcgccaaag
     2461 cccacttcac cgaccagacc gacgggatct atcaccagcc agccgcgact ggagaacatg
     2521 atgttttcat gatgcagatc gccatgtagc ccacgcagtt ccgaggcatt gctcatcatt
     2581 tgatcggcta taatcgccgc gtggacgtag tcagtttgac aacctgcgtt ttgatcatcg
     2641 cgcgcccgct gaaacaaagc tgcaaagcga tcccggatcg ggagaagggc agaaggcagg
     2701 ggttcctcag atgcggcata cagcttcgcc attagttccg ctgcaatttc ggtcgcctgg
     2761 tagtcgccgt gctcggcaac gatgtgagag agcattcgct ccccggcata ttcgagcaac
     2821 atcagattgt tctcacgacc gagcaaccgg actgctcccc tcccattgcg ccataccaga
     2881 tagtcggccc cgcgcagttc atcagcaatg tcttctatag gtttcaatcc cttgacgatt
     2941 gcaggagtcc cgtctggcaa tgaaactttc caaacgaggc tggaaaaggt gtccgcaatg
     3001 agaacaggtt gcgaaacgtg ccaatgagca ggaaaaacag gcggcatgaa catcaacccc
     3061 aagtcagagg gtccaatcgc agatagaagg caaggcgttc gcggtcgggg gcttcgatcc
     3121 ccaatacatt gaataggaca gcgaaggcgc gctctgcttc atctggcgct gcccagttct
     3181 cttcggcgtt agcaatcatg agtgccaaat cggcatagcg atctgctgtt ccgagccgcc
     3241 caaggtcgat cagacccgtg cattgaagag ttttagggtc caccatgaag ttcggcatgc
     3301 agggatcacc atggcaaaca accatatcgg tgcgctcttg gtcgagccgc accggtagct
     3361 ctcgttcgac acgagccaaa agatcgtgca gcggcgtact cttgtcctcg tccggtaaga
     3421 agtcgggatt gacggcattg cgggacacca catcaacggc gcgtccgaac attcgcgaca
     3481 gcctgcgctc aaacggacat tgatcaaccg ataggctgtg aacagcgcca agttgctgcc
     3541 ccattgacgg ccacgctttg agcaaatccg ctccagacag atcagccgcc ggtactcccg
     3601 gaattgccgt tatcaccaag catgcaccct cctgttcctc ctgccagttg atgacctcgg
     3661 ggcaagccac acctcgacct ttgagccaaa tgaggcggtc acgctctcca gcgagctcac
     3721 cgcggcggga agcaggtgcg attttcgcga aggcatgccc gtcaccacgt cgaaaaacaa
     3781 aatcaccaga ttctccgcct ctgacaggca accagtcaga atgcgattca ccaaaaaaaa
     3841 tattagttcg attcaatgga ggttccttca gttttctgat gaagcgcgaa tatagagaaa
     3901 tatcccgaat gtgcagttcc tcgagataac ttcgtatagc atacattata cgaacggtag
     3961 ctcagggcaa tgcagatccg gatgcaatag catgttctgt cagatgccat ttctgattct
     4021 tagcatcgct gcggtcttca cgggtgagct tatgctggat gaagtgccag tccggcatct
     4081 ttcttggcac agactttgct ttgtaaactg tgtcgaattg gcaccgtaaa cgcccaccat
     4141 ccttcagaag gaggtacatg gagacatccc ctttcaatat cccctgctta ggtactggta
     4201 tgatcttctc gcaggatggc tcccagttat ctgtcatctt tttcatcaca ggtccatcag
     4261 caggaaaatt cactccataa aatttggact catgatacat gcagttttct tcaacactca
     4321 ctgttatatc tgcattacat atgcaaactg ctccatcctc aaagagaaaa gacctgtccc
     4381 atgtataacc agcaggacac gagttcttga aatagtcagc tatgtcttga ggatattcag
     4441 tgaaaaccct gtttccgtac ataaaggcag ctgacaatat gtcttcggca aatggcaatg
     4501 gtccaccttc gaccacacac agattaatag cctgtttccc tttgaacgga tatccaatgc
     4561 cctctcccgt gatcacaaat ttatgtccat cgacgcaccc ttccatacgg tatttcattg
     4621 tcatttcttt tgttagaccg tgctttgact gagccatggt ggttcctcct atcgatgggg
     4681 actgacatga tcgataatga tgacatcgct ggttagaggg gaggtgccat aaaataggca
     4741 acctttacat ttcgacagaa atcatagcaa aatagtgccc ttacagatag caggaattcg
     4801 aagcttctgc agacgcgtcg acgtcatatg gatccgatat cgccgtggcg gccgctctag
     4861 aactagttag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     4921 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4981 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcaattc ttaacgacgg
     5041 taattgagaa cctgcgtaaa cggcatcagt gagttgcggg gtaagcggat gcgatattgt
     5101 tgccgcatcc ggcaaaaaaa cgggcaaggt gtcaccaccc tgcccttttt ctttaaaacc
     5161 gaaaagatta cttcgcgtga acgttttttg aagctgttat tgaaatgatt tgcagtgttt
     5221 ttgtttttcg tattttcaat gaaacaaaag caatttttta ctatctttcg cgtttcattg
     5281 attaacgact aaaatttgca ctacaggctt gcaagcccca ccaaatctaa tcccatcaga
     5341 tcccgaataa tccaatgtcc tcccggtttt tttcgtactt caagtgaatc aatacaaccc
     5401 ccattcactg ccagagccgt gagcgcagcg aactgaatgt cacgaaaaag acagcgactc
     5461 aggtgcctga tggtcggaga caaaaggaat attcagcgac tagctagtta catggctaat
     5521 tcccatgtca gccgttaagt gttcctgtgt cactcaaaat tgctttgaga ggctctaagg
     5581 gcttctcagt gcgttacatc cctggcttgt tgtccacaac cgttaaacct taaaagctag
     5641 ctttaaaagc cttatatatt cttttttttc ttataaaact taaaacctta gaggctattt
     5701 aagttgctga tttatattaa ttttattgtt caaacatgag agcttagtac gtgaaacatg
     5761 agagcttagt acgttagcca tgagagctta gtacgttagc catgagggtt tagttcgtta
     5821 aacatgagag cttagtacgt taaacatgag agcttagtac gtgaaacatg agagcttagt
     5881 acgtactatc aacaggttga actgctgatc ttcagatcct ctacgccgga cgcatcgtgg
     5941 ccggatcttg cggccgcaaa aattaaaaat gaagttttgg aggcctcatt tggtgacgaa
     6001 ataactaagc acttgtctcc tgtttactcc cctgagcttg aggggtcaac atgaaggtca
     6061 ttgatagcag gataataata cagtaaaacg ctaaaccaat aatccaaatc cagccatccc
     6121 aaattggtag tgaatgatta taaataacag taaacagtaa tgggccaata acaccggttg
     6181 cattggtaag gctcaccaat aatccctgta aagcaccttg ctcatgactc tttgtttgga
     6241 tagacatcac tccctgtaat gcaggtaaag cgatcccacc accagccaat aaaattaaaa
     6301 cagggaaatc taaccaacct tcagatataa acgctaaaaa ggcaaatgca ctactatctg
     6361 caataaattc gagcagtact gccgtttttt cgccccattt agtggctatt cttcctgcca
     6421 caaaggcttg gaatactgag tgtaaaagac caagacccgc taatgaaaag ccaaccatca
     6481 tgctattcca tccaaaacga ttttcggtaa atagcaccca caccgttgcg ggaatttggc
     6541 ctatcaattg cgctgaaaaa taaataatca acaaaatggg catcgtttta aataaagtga
     6601 tgtacaccga atttgattgc gtctcaaccc ctacttcggt atctgtatta tcacgtgtat
     6661 ttttggtttc acggaaccaa aacataacca caaggaaagt gacaatattt agcaacgcag
     6721 cgataaaaaa gggactatgc ggtgaaatct ctcctgcaaa accaccaata ataggccccg
     6781 ctattaaacc aagcccaaaa cttgccccta accaaccgaa ccacttcacg cgttgagaag
     6841 ctgaggtggt atcggcaatg accgatgccg cgacagcccc agtagctcct gtgatccctg
     6901 aaagcaaacg gcctaaatac agcatccaaa gcgcacttga aaaagccagc aataagtaat
     6961 ccagcgatgc gcctattaat gacaacaaca gcactgggcg ccgaccaaat cggtcagaca
     7021 tttttccaag ccaaggagca aagataacct gcattaacgc ataaagtgca agcaatacgc
     7081 caaagtggtt agcgatatct tccgaagcaa taaattcacg taataacgtt ggcaagactg
     7141 gcatgataag gccaatcccc atggcatcga gtaacgtaat taccaatgcg atctttgtcg
     7201 aactattcat ttcacttttc tctatcactg atagggagtg ggaaaataac tctatcaatg
     7261 atagggtgtc aa
//
