LOCUS       40924_26351        4491 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJS31, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4491)
  AUTHORS   Shen J, Chen J, Jensen PR, Solem C.
  TITLE     A novel genetic tool for metabolic optimization of Corynebacterium 
            glutamicum: efficient and repetitive chromosomal integration of 
            synthetic promoter-driven expression libraries
  JOURNAL   Appl. Microbiol. Biotechnol. (2017) In press
  PUBMED    28361238
REFERENCE   2  (bases 1 to 4491)
  AUTHORS   Shen J, Chen J, Jensen PR, Solem C.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-DEC-2016) National Food Institute, Technical 
            University of Denmark, Soltofts Plads Building 221, 056, Lyngby 
            2800, Denmark
REFERENCE   3  (bases 1 to 4491)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4491)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol. (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-DEC-2016) National Food Institute, Technical University of 
            Denmark, Soltofts Plads Building 221, 056, Lyngby 2800, Denmark"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(19..52)
                     /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)."
     regulatory      53..100
                     /label=lysA terminator from Corynebacterium glutamicum
                     /note="lysA terminator from Corynebacterium glutamicum"
                     /regulatory_class="terminator"
     gene            101..156
                     /gene="attP"
                     /label=attP
     gene            157..199
                     /gene="attB"
                     /label=attB
     misc_feature    200..414
                     /label=multiple cloning site
                     /note="multiple cloning site"
     protein_bind    complement(415..448)
                     /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)."
     rep_origin      complement(675..1263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1442..2860)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(2861..3306)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             complement(3355..4146)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 gcgtgaagct agcttatcat aacttcgtat agcatacatt atacgaacgg tacgcttttc
       61 gacgcctgac cccgcccttc accttcgccg tggagggcgg tccaactcgc ttaattgcga
      121 gtttttattt cgtttatttc aattaaggta actaaactga taattgccaa cacaattaac
      181 atctcaatca aggtaaatgc aggatcctta ggaattcaaa aaagcttccg gctgcaggac
      241 agtcgacttg acccgggaaa agcatgctac ttctagagaa tcaaactcga gaagaagtac
      301 ttggtcaatt gatttgcggc cgcatgtatg cattggggac gtcctggctt aagtaacacc
      361 ggtttaagac aaagggtcaa gtttcgaact tggaattgat tcttgcacgc gttctaccgt
      421 tcgtatagca tacattatac gaagttatta gatatcttat tggcggggag aggcggtttg
      481 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      541 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      601 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      661 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      721 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      781 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      841 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      901 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      961 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1021 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1081 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1141 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1201 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1261 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1321 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct ttttcaaggt
     1381 tgccgggcgt cgggttaccg ggcgtcgaaa agcgctttta tcattttctt ttgcgttttt
     1441 atttgttaac tgttaattgt ccttgttcaa ggatgctgtc tttgacaaca gatgttttct
     1501 tgcctttgat gttcagcagg aagctcggcg caaacgttga ttgtttgtct gcgtagaatc
     1561 ctctgtttgt catatagctt gtaatcacga cattgtttcc tttcgcttga ggtacagcga
     1621 agtgtgagta agtaaaggtt acatcgttag gatcaagatc catttttaac acaaggccag
     1681 ttttgttcag cggcttgtat gggccagtta aagaattaga aacataacca agcatgtaaa
     1741 tatcgttaga cgtaatgccg tcaatcgtca tttttgatcc gcgggagtca gtgaacaggt
     1801 accatttgcc gttcatttta aagacgttcg cgcgttcaat ttcatctgtt actgtgttag
     1861 atgcaatcag cggtttcatc acttttttca gtgtgtaatc atcgtttagc tcaatcatac
     1921 cgagagcgcc gtttgctaac tcagccgtgc gttttttatc gctttgcaga agtttttgac
     1981 tttcttgacg gaagaatgat gtgcttttgc catagtatgc tttgttaaat aaagattctt
     2041 cgccttggta gccatcttca gttccagtgt ttgcttcaaa tactaagtat ttgtggcctt
     2101 tatcttctac gtagtgagga tctctcagcg tatggttgtc gcctgagctg tagttgcctt
     2161 catcgatgaa ctgctgtaca ttttgatacg tttttccgtc accgtcaaag attgatttat
     2221 aatcctctac accgttgatg ttcaaagagc tgtctgatgc tgatacgtta acttgtgcag
     2281 ttgtcagtgt ttgtttgccg taatgtttac cggagaaatc agtgtagaat aaacggattt
     2341 ttccgtcaga tgtaaatgtg gctgaacctg accattcttg tgtttggtct tttaggatag
     2401 aatcatttgc atcgaatttg tcgctgtctt taaagacgcg gccagcgttt ttccagctgt
     2461 caatagaagt ttcgccgact ttttgataga acatgtaaat cgatgtgtca tccgcatttt
     2521 taggatctcc ggctaatgca aagacgatgt ggtagccgtg atagtttgcg acagtgccgt
     2581 cagcgttttg taatggccag ctgtcccaaa cgtccaggcc ttttgcagaa gagatatttt
     2641 taattgtgga cgaatcaaat tcagaaactt gatatttttc atttttttgc tgttcaggga
     2701 tttgcagcat atcatggcgt gtaatatggg aaatgccgta tgtttcctta tatggctttt
     2761 ggttcgtttc tttcgcaaac gcttgagttg cgcctcctgc cagcagtgcg gtagtaaagg
     2821 ttaatactgt tgcttgtttt gcaaactttt tgatgttcat cgttcatgtc tcctttttta
     2881 tgtactgtgt tagcggtctg cttcttccag ccctcctgtt tgaagatggc aagttagtta
     2941 cgcacaataa aaaaagacct aaaatatgta aggggtgacg ccaaagtata cactttgccc
     3001 tttacacatt ttaggtcttg cctgctttat cagtaacaaa cccgcgcgat ttacttttcg
     3061 acctcattct attagactct cgtttggatt gcaactggtc tattttcctc ttttgtttga
     3121 tagaaaatca taaaaggatt tgcagactac gggcctaaag aactaaaaaa tctatctgtt
     3181 tcttttcatt ctctgtattt tttatagttt ctgttgcatg ggcataaagt tgccttttta
     3241 atcacaattc agaaaatatc ataatatctc atttcactaa ataatagtga acggcaggta
     3301 tatgtgatgg gttaaaaagg atcgatcctc tagcgaaccc cagagtcccg ctcagaagaa
     3361 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     3421 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     3481 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     3541 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     3601 ctcgccgtcg ggcatccgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     3661 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     3721 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     3781 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     3841 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     3901 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     3961 gtcttggagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     4021 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     4081 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     4141 aatcatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca
     4201 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga
     4261 gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta
     4321 tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt
     4381 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     4441 ctacgtgttc cgcttccttt agcagccctt gcgccctgag tgcttgcggc a
//
