LOCUS       V001868                 5315 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V001868
VERSION     V001868
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5315)
  AUTHORS   Shenkerman Y, Elharar Y, Vishkautzan M, Gur E.
  TITLE     Efficient and simple generation of unmarked gene deletions in
            Mycobacterium smegmatis
  JOURNAL   Gene 533 (1), 374-378 (2014)
   PUBMED   24100088
REFERENCE   2  (bases 1 to 5315)
  AUTHORS   Shenkerman Y, Elharar Y, Vishkautzan M, Gur E.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-AUG-2013) Life Sciences, Ben-Gurion University of the
            Negev, Rager Blvd, Beer-Sheva 84105, Israel
REFERENCE   3  (bases 1 to 5315)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5315)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
            date: "2014"; volume: "533"; issue: "1"; pages: "374-378"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-AUG-2013) Life Sciences, Ben-Gurion University of the Negev,
            Rager Blvd, Beer-Sheva 84105, Israel"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    616..649
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1008..1721
                     /label="mgfp5"
                     /note="GFP with folding enhancement mutations"
     CDS             1836..2831
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     misc_recomb     3059..3092
                     /label="loxP site for cre recombinase"
                     /note="loxP site for cre recombinase"
     protein_bind    complement(3059..3092)
                     /label="loxP"
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     promoter        3409..3513
                     /label="AmpR promoter"
     CDS             3514..4371
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4545..5133
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg aattcgagct
      241 cggtacccgg ggatcctcgg tcgacgtgtg cgcgtagccg ctgcgccaga acacccgcat
      301 cgcggtgtcc agtgcgacgt ccggatcgaa ctccttgtga cgggccatgt gccaattcta
      361 gatcgtcgag aaggtcgacg gccagtggag gacgtactgc tccgagacct gctactggac
      421 cgacgccgtc gcgttccggg gtgagtacga gggcagggaa accccgaaca tgggacggct
      481 caccgggttc cgagaatggg agaccctgca ccacggcaat ccaatattga tccactagtt
      541 taaaccaaga agatcctttg atattttcta cggggtctga cgctcagtcg agtatacgag
      601 gcgcgccaga tttaaataac ttcgtataat gtatgctata cgaagttatc ccagatctgt
      661 aagataaact ctagaaatat tggatcgtcg gcaccgtcac ggccgtggga ggcggcacga
      721 tccgcgacgt gatgatcggc cgcatcccca cggtgctgcg cagtgagctc tacgccatcc
      781 cggcgttgat ctgtgcgttc gcacgcacag gcccggtgtg agaagggtct ctgacgagcg
      841 ggagaaccca cccggggtgg gcgagtttgt cctgcgtgtg ctcggtcgag taggctctgg
      901 gagtacccgt gtgtacgacc agcacggcat acatcatttc gacgccgaga gattcgccgc
      961 ccgaaatgag cacgatccgc atgcttataa cagaaaggag gttaataatg tcgaagggcg
     1021 aggagctgtt caccggcgtc gtcccgatcc tggtcgagct ggacggtgac gtcaacggcc
     1081 acaagttctc cgtctccggc gagggtgagg gcgacgccac ctacggcaag ctgaccctga
     1141 agttcatctg caccaccggt aagctgccgg tcccgtggcc gaccctggtc accaccctga
     1201 cctacggcgt ccagtgcttc tcccgctacc cggaccacat gaagcgccac gacttcttca
     1261 agtccgccat gccggagggt tacgtccagg agcgcaccat ctccttcaag gacgacggta
     1321 actacaagac gcgtgccgag gtcaagttcg agggcgacac cctggtcaac cgcatcgagc
     1381 tgaagggcat cgacttcaag gaggacggta acatcctggg ccacaagctg gagtacaact
     1441 acaactccca caacgtctac atcaccgcgg acaagcagaa gaacggcatc aaggccaact
     1501 tcaagacccg ccacaacatc gaggacggtg gcgtccagct agccgaccac taccagcaga
     1561 acaccccgat cggcgacggc ccggtcctgc tgccggacaa ccactacctg tccacccagt
     1621 ccgccctgtc caaggacccg aacgagaagc gcgaccacat ggtcctgctg gagttcgtca
     1681 ccgccgccgg catcacccac ggcatggacg agctgtacaa gtagatttat caccagcccg
     1741 tcatcgtcaa cgcctgatcc gcggtgcgga caggccgtgt cgcgaccggc cgtgcggaat
     1801 taagccggcc cgtaccctgt gaatagaggt ccgctgtgac acaagaatcc ctgttacttc
     1861 tcgaccgtat tgattcggat gattcctacg cgagcctgcg gaacgaccag gaattctggg
     1921 agccgctggc ccgccgagcc ctggaggagc tcgggctgcc ggtgccgccg gtgctgcggg
     1981 tgcccggcga gagcaccaac cccgtactgg tcggcgagcc cgacccggtg atcaagctgt
     2041 tcggcgagca ctggtgcggt ccggagagcc tcgcgtcgga gtcggaggcg tacgcggtcc
     2101 tggcggacgc cccggtgccg gtgccccgcc tcctcggccg cggcgagctg cggcccggca
     2161 ccggagcctg gccgtggccc tacctggtga tgagccggat gaccggcacc acctggcggt
     2221 ccgcgatgga cggcacgacc gaccggaacg cgctgctcgc cctggcccgc gaactcggcc
     2281 gggtgctcgg ccggctgcac agggtgccgc tgaccgggaa caccgtgctc accccccatt
     2341 ccgaggtctt cccggaactg ctgcgggaac gccgcgcggc gaccgtcgag gaccaccgcg
     2401 ggtggggcta cctctcgccc cggctgctgg accgcctgga ggactggctg ccggacgtgg
     2461 acacgctgct ggccggccgc gaaccccggt tcgtccacgg cgacctgcac gggaccaaca
     2521 tcttcgtgga cctggccgcg accgaggtca ccgggatcgt cgacttcacc gacgtctatg
     2581 cgggagactc ccgctacagc ctggtgcaac tgcatctcaa cgccttccgg ggcgaccgcg
     2641 agatcctggc cgcgctgctc gacggggcgc agtggaagcg gaccgaggac ttcgcccgcg
     2701 aactgctcgc cttcaccttc ctgcacgact tcgaggtgtt cgaggagacc ccgctggatc
     2761 tctccggctt caccgatccg gaggaactgg cgcagttcct ctgggggccg ccggacaccg
     2821 cccccggcgc ctgacgcccc gggccgcccg gcgccgcccc cggcccccgg cggccatcga
     2881 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
     2941 gttcggtaaa ttgtcacaac gaattcgata aaataaaaaa ggggacctct agggtcccct
     3001 tttttatttt atcgaattcg agctcgaagt tcctattctc tagactgcag gaacgcgtat
     3061 aacttcgtat aatgtatgct atacgaagtt attaattaaa gctagcacat gcatggttat
     3121 tgtctcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3181 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3241 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3301 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     3361 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3421 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3481 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3541 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3601 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3661 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3721 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3781 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3841 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3901 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3961 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4021 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4081 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4141 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4201 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4261 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4321 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4381 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4441 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4501 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4561 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4621 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4681 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4741 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4801 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4861 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4921 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4981 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5041 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5101 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5161 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5221 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5281 gagcgcagcg agtcagtgag cgaggaagcg gaaga
//
