LOCUS       62056_13735        7938 bp DNA     circular SYN 13-MAY-2019
DEFINITION  Cloning vector pLCB1, complete sequence.
ACCESSION   MK176919
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7938)
  AUTHORS   Li L.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-NOV-2018) Key Laboratory of Synthetic Biology, 
            Institute of Plant Physiology and Ecology, 300 Feng Lin Road, 
            Shanghai, Shanghai 200032, China
REFERENCE   2  (bases 1 to 7938)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (13-NOV-2018) Key Laboratory of Synthetic Biology, Institute of 
            Plant Physiology and Ecology, 300 Feng Lin Road, Shanghai, Shanghai 
            200032, China"
FEATURES             Location/Qualifiers
     source          1..7938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             220..2004
                     /codon_start=1
                     /transl_table=11
                     /product="phiBT1 integrase"
                     /label=phiBT1 integrase
                     /protein_id="QCO69769.1"
                     /translation="MSPFIAPDVPEHLLDTVRVFLYARQSKGRSDGSDVSTEAQLAAGR
                     ALVASRNAQGGARWVVAGEFVDVGRSGWDPNVTRADFERMMGEVRAGEGDVVVVNELSR
                     LTRKGAHDALEIDNELKKHGVRFMSVLEPFLDTSTPIGVAIFALIAALAKQDSDLKAER
                     LKGAKDEIAALGGVHSSSAPFGMRAVRKKVDNLVISVLEPDEDNPDHVELVERMAKMSF
                     EGVSDNAIATTFEKEKIPSPGMAERRATEKRLASIKARRLNGAEKPIMWRAQTVRWILN
                     HPAIGGFAFERVKHGKAHINVIRRDPGGKPLTPHTGILSGSKWLELQEKRSGKNLSDRK
                     PGAEVEPTLLSGWRFLGCRICGGSMGQSQGGRKRNGDLAEGNYMCANPKGHGGLSVKRS
                     ELDEFVASKVWARLRTADMEDEHDQAWIAAAAERFALQHDLAGVADERREQQAHLDNVR
                     RSIKDLQADRKAGLYVGREELETWRSTVLQYRSYEAECTTRLAELDEKMNGSTRVPSEW
                     FSGEDPTAEGGIWASWDVYERREFLSFFLDSVMVDRGRHPETKKYIPLKDRVTLKWAEL
                     LKEEDEASEATERELAAL"
     primer_bind     2499..2515
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2565..2661)
                     /regulatory_class="promoter"
     primer_bind     complement(2682..2698)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2706..2722)
                     /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)."
     promoter        complement(2730..2760)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2775..2796)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3084..3672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3925..4725
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     CDS             complement(5162..5530)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(5563..5672)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(5910..5921)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     protein_bind    5992..6091
                     /label=phage phi-C31 attP
                     /note="attachment site of phage phi-C31"
     CDS             6101..7915
                     /label=phage phi-C31 integrase
                     /note="integrase from phage phi-C31"
ORIGIN
        1 atctacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
       61 gctctcgcag ggcgaagttt aaactggcgc cggacggggc ttcagacgtt tcgggtgctg
      121 ggttgttgtc tctggacagt gatccatggg aaactactca gcaccaccaa tgttcccaaa
      181 agaaagcgca ggtcagcgcc catgagccaa gatctaggca tgtcgccctt catcgctccc
      241 gacgtccctg agcaccttct agacactgtt cgcgtcttcc tgtacgcgcg tcagtctaag
      301 ggccggtccg acggctcaga cgtgtcgacc gaagcacagc tagcggccgg tcgtgcgttg
      361 gtcgcgtctc gcaacgccca ggggggtgcg cgctgggtcg tggcaggtga gttcgtggac
      421 gtcgggcgct ccggctggga cccgaacgtg acccgtgccg acttcgagcg catgatgggc
      481 gaagtccgcg ccggcgaagg tgacgttgtc gttgtgaatg agctttcccg gctcactcgc
      541 aagggcgccc atgacgcgct cgaaatcgac aacgaattga agaagcacgg cgtgcgcttc
      601 atgtcggttc ttgagccgtt ccttgacacg tctaccccta tcggcgtcgc cattttcgcg
      661 ctgatcgctg cccttgcgaa acaggacagt gacctgaagg cggagcgcct gaagggtgcg
      721 aaagacgaga ttgccgcgct gggtggcgtt cactcgtctt ccgccccgtt cggaatgcgc
      781 gccgtgcgca agaaggtcga taatctcgtg atctccgttc ttgagccgga cgaagacaac
      841 ccggatcacg tcgagctagt tgagcgcatg gcgaaaatgt cgttcgaagg cgtgtccgac
      901 aacgccattg caacgacctt cgagaaggaa aagatcccgt cgcccggaat ggctgagaga
      961 cgcgccacgg aaaagcgtct tgcgtccatc aaggcacgtc gcctgaacgg cgctgaaaag
     1021 ccgatcatgt ggcgcgctca aacggtccga tggattctca accatcccgc aatcggcggt
     1081 ttcgcattcg agcgtgtgaa gcacggtaag gcgcacatca acgtcatacg gcgcgacccc
     1141 ggcggcaagc cgctaacgcc ccacacgggc attctcagcg gctcgaagtg gcttgagctt
     1201 caagagaagc gttccgggaa gaatctcagc gaccggaagc ctggggccga agtcgaaccg
     1261 acgcttctga gcgggtggcg tttcctgggg tgccgaatct gcggcggctc aatgggtcag
     1321 tcccagggtg gccgtaagcg caacggcgac cttgccgaag gcaattacat gtgcgccaac
     1381 ccgaaggggc acggcggctt gtcggtcaag cgcagcgaac tggacgaatt cgttgcttcg
     1441 aaggtgtggg cacggctccg cacagccgac atggaagatg aacacgatca ggcatggatt
     1501 gccgccgctg cggagcgctt cgcccttcag cacgacctag cgggggtggc cgatgagcgg
     1561 cgcgaacaac aggcgcacct agacaacgtg cggcgctcca tcaaggacct tcaggcggac
     1621 cgtaaggccg gtctgtacgt cgggcgtgaa gagctggaaa cgtggcgctc aacggtgctg
     1681 caataccggt cctacgaagc ggagtgcacg acccgactcg ctgagcttga cgagaagatg
     1741 aacggcagca cccgcgttcc gtctgagtgg ttcagcggcg aagacccgac ggccgaaggg
     1801 ggcatctggg caagctggga cgtgtacgag cgtcgggagt tcctgagctt cttccttgac
     1861 tccgtcatgg tcgaccgggg gcgccaccct gagacgaaga aatacatccc cctgaaggac
     1921 cgtgtgacgc tcaagtgggc ggagctgctg aaggaggaag acgaagcgag cgaagccact
     1981 gagcgggagc ttgcggcgct gtagcgcaca gcgggagggg tcgagccggc ggacggttcg
     2041 gccccttttt tggccttgaa atcgttagtt aggctaatgc atatgcggcc gccgcaattg
     2101 atttaaattg aagccgccga agccccgaag cttccccttg accaatacta gtgaatctcg
     2161 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     2221 tggtttctac aaagatcgtt atgttgatcg gcactttgca tcggccgcgc tcccgattcc
     2281 ggaagtgctt gacattgggg aatttatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2341 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2401 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2461 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     2521 gcttgggctg caggtcgact ctagaggatc cttaattaag gcatatggac actccttact
     2581 tagactgtcg tattctcctg gccacgactt tacaacaccg cacagcatgt tgtcaaagca
     2641 gagacggttc gaatgtgaac agatatcgaa ttcgtaatca tgtcatagct gtttcctgtg
     2701 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2761 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2821 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2881 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3001 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3481 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3661 aaaggatctc aagaagatcc tttgatctac tagtagctcc atcagcaaaa ggggatgata
     3721 agtttatcac caatttaaat ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3781 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tggttcatgt
     3841 gcagctccat cagcaaaagg ggatgataag tttatcacca ccgactattt gcaacagtgc
     3901 cgttgatcgt gctatgatcg actgatgtca tcagcggtgg agtgcaatgt cgtgcaatac
     3961 gaatggcgaa aagccgagct catcggtcag cttctcaacc ttggggttac ccccggcggt
     4021 gtgctgctgg tccacagctc cttccgtagc gtccggcccc tcgaagatgg gccacttgga
     4081 ctgatcgagg ccctgcgtgc tgcgctgggt ccgggaggga cgctcgtcat gccctcgtgg
     4141 tcaggtctgg acgacgagcc gttcgatcct gccacgtcgc ccgttacacc ggaccttgga
     4201 gttgtctctg acacattctg gcgcctgcca aatgtaaagc gcagcgccca tccatttgcc
     4261 tttgcggcag cggggccaca ggcagagcag atcatctctg atccattgcc cctgccacct
     4321 cactcgcctg caagcccggt cgcccgtgtc catgaactcg atgggcaggt acttctcctc
     4381 ggcgtgggac acgatgccaa cacgacgctg catcttgccg agttgatggc aaaggttccc
     4441 tatggggtgc cgagacactg caccattctt caggatggca agttggtacg cgtcgattat
     4501 ctcgagaatg accactgctg tgagcgcttt gccttggcgg acaggtggct caaggagaag
     4561 agccttcaga aggaaggtcc agtcggtcat gcctttgctc ggttgatccg ctcccgcgac
     4621 attgtggcga cagccctggg tcaactgggc cgagatccgt tgatcttcct gcatccgcca
     4681 gaggcgggat gcgaagaatg cgatgccgct cgccagtcga ttggctgagc tcatgagcgg
     4741 agaacgagat gacgttggag gggcaaggtc gcgctgattg ctggggcaac acgtggagcg
     4801 gatcggggat tgtctttctt cagctcgctg atgatatgct gacgctcaat gccgtttggc
     4861 ctccgactaa cgaaaatccc gcatttggac ggctgatccg attggcacgg cggacggcga
     4921 atggcggagc agacgctcgt ccgggggcaa tgagatatga aaaagcctga actcaccgcg
     4981 acgtatcggg ccctggccag ctagctagag tcgacctgca ggtccccggg gatcggtctt
     5041 gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct tgatggagcg
     5101 catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc
     5161 atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct
     5221 cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt
     5281 cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg
     5341 ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg
     5401 ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc
     5461 gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt
     5521 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     5581 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     5641 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     5701 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     5761 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagatc cgtcgacctg
     5821 caggcatgca agctctagcg attccagacg tcccgaaggc gtggcgcggc ttccccgtgc
     5881 cggagcaatc gccctgggtg ggttacacga cgcccctcta tggcccgtac tgacggacac
     5941 accgaagccc cggcggcaac cctcagcgga tgccccgggg cttcacgttt tcccaggtca
     6001 gaagcggttt tcgggagtag tgccccaact ggggtaacct ttgagttctc tcagttgggg
     6061 gcgtagggtc gccgacatga cacaaggggt tgtgaccggg gtggacacgt acgcgggtgc
     6121 ttacgaccgt cagtcgcgcg agcgcgaaaa ttcgagcgca gcaagcccag cgacacagcg
     6181 tagcgccaac gaagacaagg cggccgacct tcagcgcgaa gtcgagcgcg acgggggccg
     6241 gttcaggttc gtcgggcatt tcagcgaagc gccgggcacg tcggcgttcg ggacggcgga
     6301 gcgcccggag ttcgaacgca tcctgaacga atgccgcgcc gggcggctca acatgatcat
     6361 tgtctatgac gtgtcgcgct tctcgcgcct gaaggtcatg gacgcgattc cgattgtctc
     6421 ggaattgctc gccctgggcg tgacgattgt ttccactcag gaaggcgtct tccggcaggg
     6481 aaacgtcatg gacctgattc acctgattat gcggctcgac gcgtcgcaca aagaatcttc
     6541 gctgaagtcg gcgaagattc tcgacacgaa gaaccttcag cgcgaattgg gcgggtacgt
     6601 cggcgggaag gcgccttacg gcttcgagct tgtttcggag acgaaggaga tcacgcgcaa
     6661 cggccgaatg gtcaatgtcg tcatcaacaa gcttgcgcac tcgaccactc cccttaccgg
     6721 acccttcgag ttcgagcccg acgtaatccg gtggtggtgg cgtgagatca agacgcacaa
     6781 acaccttccc ttcaagccgg gcagtcaagc cgccattcac ccgggcagca tcacggggct
     6841 ttgtaagcgc atggacgctg acgccgtgcc gacccggggc gagacgattg ggaagaagac
     6901 cgcttcaagc gcctgggacc cggcaaccgt tatgcgaatc cttcgggacc cgcgtattgc
     6961 gggcttcgcc gctgaggtga tctacaagaa gaagccggac ggcacgccga ccacgaagat
     7021 tgagggttac cgcattcagc gcgacccgat cacgctccgg ccggtcgagc ttgattgcgg
     7081 accgatcatc gagcccgctg agtggtatga gcttcaggcg tggttggacg gcagggggcg
     7141 cggcaagggg ctttcccggg ggcaagccat tctgtccgcc atggacaagc tgtactgcga
     7201 gtgtggcgcc gtcatgactt cgaagcgcgg ggaagaatcg atcaaggact cttaccgctg
     7261 ccgtcgccgg aaggtggtcg acccgtccgc acctgggcag cacgaaggca cgtgcaacgt
     7321 cagcatggcg gcactcgaca agttcgttgc ggaacgcatc ttcaacaaga tcaggcacgc
     7381 cgaaggcgac gaagagacgt tggcgcttct gtgggaagcc gcccgacgct tcggcaagct
     7441 cactgaggcg cctgagaaga gcggcgaacg ggcgaacctt gttgcggagc gcgccgacgc
     7501 cctgaacgcc cttgaagagc tgtacgaaga ccgcgcggca ggcgcgtacg acggacccgt
     7561 tggcaggaag cacttccgga agcaacaggc agcgctgacg ctccggcagc aaggggcgga
     7621 agagcggctt gccgaacttg aagccgccga agccccgaag cttccccttg accaatggtt
     7681 ccccgaagac gccgacgctg acccgaccgg ccctaagtcg tggtgggggc gcgcgtcagt
     7741 agacgacaag cgcgtgttcg tcgggctctt cgtagacaag atcgttgtca cgaagtcgac
     7801 tacgggcagg gggcagggaa cgcccatcga gaagcgcgct tcgatcacgt gggcgaagcc
     7861 gccgaccgac gacgacgaag acgacgccca ggacggcacg gaagacgtag cggcgtagcg
     7921 agacacccgg gaagcctg
//
