LOCUS       40924_2438        7277 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p17YFP(lox-Sm-E-Kn)-Tc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  AUTHORS   Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV.
  TITLE     Genetic tools for C. glutamicum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7277)
  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 7277)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7277)
  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            "Unpublished"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..7277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  complement(58..175)
                     /mobile_element_type="transposon:Mu bacteriophage attR"
                     /label=ttR
     protein_bind    complement(282..315)
                     /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    361..394
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(588..1379)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    1740..1787
                     /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(1993..2140)
                     /regulatory_class="enhancer"
     CDS             complement(2219..3055)
                     /codon_start=1
                     /gene="strB"
                     /product="StrB"
                     /label=strB
                     /note="streptomycin phosphotransferase"
                     /protein_id="AKN19590.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     gene            complement(2219..3055)
                     /gene="strB"
                     /label=strB
     CDS             complement(3055..3858)
                     /codon_start=1
                     /gene="strB"
                     /product="StrA"
                     /label=strB
                     /note="streptomycin phosphotransferase"
                     /protein_id="AKN19591.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     gene            complement(3055..3858)
                     /gene="strB"
                     /label=strB
     protein_bind    complement(3928..3961)
                     /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..4678)
                     /label=Citrine
                     /note="enhanced variant of YFP (Heikal et al., 2001)"
     primer_bind     complement(4893..4909)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5112..5171
                     /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(5183..5400)
                     /mobile_element_type="transposon:Mu bacteriophage attL"
                     /label=ttL
     rep_origin      complement(5530..5922)
                     /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(6012..7214)
                     /label=TcR
                     /note="tetracycline efflux protein"
     protein_bind    complement(7223..7241)
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 tgcatgatag tttccttgca tgaatccata aaaggcgcct gtagtgccat ttacccctga
       61 agcggcgcac gaaaaacgcg aaagcgtttc acgataaatg cgaaaacttt agctttcgcg
      121 cttcaaatga aacagatgta ttaattactg ctttttattc attacatggg gatcttgaaa
      181 gctggagaga aaaatgaata gatactaaaa agccggagca gtctcccgct ccggcttcac
      241 aaggcaatcg ccttgcagcg aaacacaatt actcttatcg ctaccgttcg tatagcatac
      301 attatacgaa gttatgattc cagtgcgatt ttgatcatct gcatccttag ttcctattcc
      361 gaagttccta ttctctagaa agtataggaa cttcagagcg cttttgaagc tggggtgggc
      421 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
      481 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
      541 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
      601 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
      661 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
      721 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
      781 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
      841 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
      901 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
      961 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     1021 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     1081 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     1141 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     1201 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     1261 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     1321 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     1381 cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt
     1441 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     1501 ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat
     1561 gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata
     1621 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg
     1681 ttccgcttcc tttagcagcc cttgcgccct gagtgcttgc ggcagcgtga ggggatcttg
     1741 aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcgaa gcagctccag
     1801 cctacacaat cgctcaagac gtgtaatgct gcaagtcgac catcgacatc acatcgtatt
     1861 caacagcttt cccccccttg acaccttcct ttttttgctt cgtccagcct tgaacatttg
     1921 ctcgatagtg aacaccagca acagatccag gcataccatc agcagccata atttcttgcg
     1981 gcgaacacca aattgacttt tcagtattat tcttttctat aaagttactt ttcaaaattt
     2041 aaactcctta tttatcaacg cgttaatcag taatcaaagg aatttaccaa aaagcagctt
     2101 tacaaaaagc ttttcagtaa ttatcttttt agtaagctag ctaagttttt acacttagtt
     2161 aaattgctaa ctttatagat tacaaaactt aggagggttt ttaaatgtgg atccccatct
     2221 agtatgacgt ctgtcgcacc tgcttgatcg cggccgcgat agctagatcg cgttgctcct
     2281 cttctccatc cgcgttccaa gctgcggaaa ggcacccata agcgtacgcc tggtcgagca
     2341 ggcgacgcgg atcgacgtcc agcgcacgag agaatgcgtc cgccatctgt gcaatgcgtc
     2401 taggatcgag acaaaggtcg tctctgtcag ccggatcgta gaacatattg gcggcgccaa
     2461 agcccacttc accgaccaga ccgacgggat ctatcaccag ccagccgcga ctggagaaca
     2521 tgatgttttc atgatgcaga tcgccatgta gcccacgcag ttccgaggca ttgctcatca
     2581 tttgatcggc tataatcgcc gcgtggacgt agtcagtttg acaacctgcg ttttgatcat
     2641 cgcgcgcccg ctgaaacaaa gctgcaaagc gatcccggat cgggagaagg gcagaaggca
     2701 ggggttcctc agatgcggca tacagcttcg ccattagttc cgctgcaatt tcggtcgcct
     2761 ggtagtcgcc gtgctcggca acgatgtgag agagcattcg ctccccggca tattcgagca
     2821 acatcagatt gttctcacga ccgagcaacc ggactgctcc cctcccattg cgccatacca
     2881 gatagtcggc cccgcgcagt tcatcagcaa tgtcttctat aggtttcaat cccttgacga
     2941 ttgcaggagt cccgtctggc aatgaaactt tccaaacgag gctggaaaag gtgtccgcaa
     3001 tgagaacagg ttgcgaaacg tgccaatgag caggaaaaac aggcggcatg aacatcaacc
     3061 ccaagtcaga gggtccaatc gcagatagaa ggcaaggcgt tcgcggtcgg gggcttcgat
     3121 ccccaataca ttgaatagga cagcgaaggc gcgctctgct tcatctggcg ctgcccagtt
     3181 ctcttcggcg ttagcaatca tgagtgccaa atcggcatag cgatctgctg ttccgagccg
     3241 cccaaggtcg atcagacccg tgcattgaag agttttaggg tccaccatga agttcggcat
     3301 gcagggatca ccatggcaaa caaccatatc ggtgcgctct tggtcgagcc gcaccggtag
     3361 ctctcgttcg acacgagcca aaagatcgtg cagcggcgta ctcttgtcct cgtccggtaa
     3421 gaagtcggga ttgacggcat tgcgggacac cacatcaacg gcgcgtccga acattcgcga
     3481 cagcctgcgc tcaaacggac attgatcaac cgataggctg tgaacagcgc caagttgctg
     3541 ccccattgac ggccacgctt tgagcaaatc cgctccagac agatcagccg ccggtactcc
     3601 cggaattgcc gttatcacca agcatgcacc ctcctgttcc tcctgccagt tgatgacctc
     3661 ggggcaagcc acacctcgac ctttgagcca aatgaggcgg tcacgctctc cagcgagctc
     3721 accgcggcgg gaagcaggtg cgattttcgc gaaggcatgc ccgtcaccac gtcgaaaaac
     3781 aaaatcacca gattctccgc ctctgacagg caaccagtca gaatgcgatt caccaaaaaa
     3841 aatattagtt cgattcaatg gaggttcctt cagttttctg atgaagcgcg aatatagaga
     3901 aatatcccga atgtgcagtt cctcgagata acttcgtata gcatacatta tacgaacggt
     3961 attatttgta caattcatca ataccatggg taataccagc agcagtaaca aattctaaca
     4021 agaccatgtg gtctctcttt tcgtttggat ctttggataa ggcagattga taggataagt
     4081 aatggttgtc tggtaacaag actggaccat caccaattgg agtattttgt tgataatggt
     4141 cagctaattg aacagaacca tcttcaatgt tgtgtctaat tttgaagtta actttgatac
     4201 cattcttttg tttgtcagcc atgatgtaaa cattgtgaga gttatagttg tattccaatt
     4261 tgtgacctaa aatgttacca tcttctttaa aatcaatacc ttttaattcg attctattaa
     4321 ctaaggtatc accttcaaac ttgacttcag ctctggtctt gtagttaccg tcatctttga
     4381 aaaaaatagt tctttcttga acataacctt ctggcatggc agacttgaaa aagtcatgtt
     4441 gtttcatatg atctgggtat ctagcaaaac acatcaaacc ataacctaaa gtagtgacta
     4501 aggttggcca tggaactggc aatttaccag tagtacaaat aaattttaag gtcaatttac
     4561 cgtaagtagc atcaccttca ccttcaccgg agacagaaaa tttgtgacca ttaacatcac
     4621 catctaattc aaccaaaatt gggacaacac cagtgaataa ttcttcacct ttagacatgt
     4681 taattaacct cctatcgata atgatgacat cgctggttag aggggaggtg ccataaaata
     4741 ggcaaccttt acatttcgac agaaatcata gcaaaatagt gcccttacag atagcaggaa
     4801 ttcgaagctt ctgcagacgc gtcgacgtca tatggatccg atatcgccgt ggcggccgct
     4861 ctagaactag ttagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
     4921 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4981 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgca attcttaacg
     5041 acggtaattg agaacctgcg taaacggcat cagtgagttg cggggtaagc ggatgcgata
     5101 ttgttgccgc atccggcaaa aaaacgggca aggtgtcacc accctgccct ttttctttaa
     5161 aaccgaaaag attacttcgc gtgaacgttt tttgaagctg ttattgaaat gatttgcagt
     5221 gtttttgttt ttcgtatttt caatgaaaca aaagcaattt tttactatct ttcgcgtttc
     5281 attgattaac gactaaaatt tgcactacag gcttgcaagc cccaccaaat ctaatcccat
     5341 cagatcccga ataatccaat gtcctcccgg tttttttcgt acttcaagtg aatcaataca
     5401 acccccattc actgccagag ccgtgagcgc agcgaactga atgtcacgaa aaagacagcg
     5461 actcaggtgc ctgatggtcg gagacaaaag gaatattcag cgactagcta gttacatggc
     5521 taattcccat gtcagccgtt aagtgttcct gtgtcactga aaattgcttt gagaggctct
     5581 aagggcttct cagtgcgtta catccctggc ttgttgtcca caaccgttaa accttaaaag
     5641 ctagctttaa aagccttata tattcttttt tttcttataa aacttaaaac cttagaggct
     5701 atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta gtacgtgaaa
     5761 catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag ggtttagttc
     5821 gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa catgagagct
     5881 tagtacgtac tatcaacagg ttgaactgct gatcttcaga tcctctacgc cggacgcatc
     5941 gtggccggat cttgcggccg caaaaattaa aaatgaagtt ttggaggcct catttggtga
     6001 cgaaataact aagcacttgt ctcctgttta ctcccctgag cttgaggggt caacatgaag
     6061 gtcattgata gcaggataat aatacagtaa aacgctaaac caataatcca aatccagcca
     6121 tcccaaattg gtagtgaatg attataaata acagtaaaca gtaatgggcc aataacaccg
     6181 gttgcattgg taaggctcac caataatccc tgtaaagcac cttgctcatg actctttgtt
     6241 tggatagaca tcactccctg taatgcaggt aaagcgatcc caccaccagc caataaaatt
     6301 aaaacaggga aatctaacca accttcagat ataaacgcta aaaaggcaaa tgcactacta
     6361 tctgcaataa attcgagcag tactgccgtt ttttcgcccc atttagtggc tattcttcct
     6421 gccacaaagg cttggaatac tgagtgtaaa agaccaagac ccgctaatga aaagccaacc
     6481 atcatgctat tccatccaaa acgattttcg gtaaatagca cccacaccgt tgcgggaatt
     6541 tggcctatca attgcgctga aaaataaata atcaacaaaa tgggcatcgt tttaaataaa
     6601 gtgatgtaca ccgaatttga ttgcgtctca acccctactt cggtatctgt attatcacgt
     6661 gtatttttgg tttcacggaa ccaaaacata accacaagga aagtgacaat atttagcaac
     6721 gcagcgataa aaaagggact atgcggtgaa atctctcctg caaaaccacc aataataggc
     6781 cccgctatta aaccaagccc aaaacttgcc cctaaccaac cgaaccactt cacgcgttga
     6841 gaagctgagg tggtatcggc aatgaccgat gccgcgacag ccccagtagc tcctgtgatc
     6901 cctgaaagca aacggcctaa atacagcatc caaagcgcac ttgaaaaagc cagcaataag
     6961 taatccagcg atgcgcctat taatgacaac aacagcactg ggcgccgacc aaatcggtca
     7021 gacatttttc caagccaagg agcaaagata acctgcatta acgcataaag tgcaagcaat
     7081 acgccaaagt ggttagcgat atcttccgaa gcaataaatt cacgtaataa cgttggcaag
     7141 actggcatga taaggccaat ccccatggca tcgagtaacg taattaccaa tgcgatcttt
     7201 gtcgaactat tcatttcact tttctctatc actgataggg agtgggaaaa taactctatc
     7261 aatgataggg tgtcacc
//
