LOCUS       62056_7980        5214 bp DNA     circular SYN 17-MAY-2021
DEFINITION  Cloning vector pCrRNA-V, complete sequence.
ACCESSION   MW285724
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5214)
  AUTHORS   Lin Z, Li H, He L, Jing Y, Pistolozzi M, Wang T, Ye Y.
  TITLE     Efficient genome editing for Pseudomonas aeruginosa using 
            CRISPR-Cas12a
  JOURNAL   Gene, 145693 (2021) In press
  PUBMED    33961975
REFERENCE   2  (bases 1 to 5214)
  AUTHORS   Lin Z, Li H.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-NOV-2020) School of Biology and Biological 
            Engineering, South China University of Technology, 382 East Outer 
            Loop Road, University Park, Guangzhou, Guangdong 510006, China
REFERENCE   3  (bases 1 to 5214)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene, 
            145693 (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-NOV-2020) School of Biology and Biological Engineering, South 
            China University of Technology, 382 East Outer Loop Road, University
            Park, Guangzhou, Guangdong 510006, China"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..301
                     /regulatory_class="promoter"
     CDS             302..1720
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     terminator      1747..1841
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1872..1900
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2089..2619
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     oriT            2833..2941
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      3035..3623
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3680..4510)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      4524..4875
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     terminator      complement(4948..5034)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        5071..5105
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological
                     Parts BBa_J23119)"
     misc_feature    5106..5124
                     /label=5' handle with the crRNA
                     /note="5' handle with the crRNA"
     regulatory      5147..5153
                     /regulatory_class="terminator"
ORIGIN
        1 tctttaggcc cgtagtctgc aaatcctttt atgattttct atcaaacaaa agaggaaaat
       61 agaccagttg caatccaaac gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg
      121 gtttgttact gataaagcag gcaagaccta aaatgtgtaa agggcaaagt gtatactttg
      181 gcgtcacccc ttacatattt taggtctttt tttattgtgc gtaactaact tgccatcttc
      241 aaacaggagg gctggaagaa gcagaccgct aacacagtac ataaaaaagg agacatgaac
      301 gatgaacatc aaaaagtttg caaaacaagc aacagtatta acctttacta ccgcactgct
      361 ggcaggaggc gcaactcaag cgtttgcgaa agaaacgaac caaaagccat ataaggaaac
      421 atacggcatt tcccatatta cacgccatga tatgctgcaa atccctgaac agcaaaaaaa
      481 tgaaaaatat caagttcctg aattcgattc gtccacaatt aaaaatatct cttctgcaaa
      541 aggcctggac gtttgggaca gctggccatt acaaaacgct gacggcactg tcgcaaacta
      601 tcacggctac cacatcgtct ttgcattagc cggagatcct aaaaatgcgg atgacacatc
      661 gatttacatg ttctatcaaa aagtcggcga aacttctatt gacagctgga aaaacgctgg
      721 ccgcgtcttt aaagacagcg acaaattcga tgcaaatgat tctatcctaa aagaccaaac
      781 acaagaatgg tcaggttcag ccacatttac atctgacgga aaaatccgtt tattctacac
      841 tgatttctcc ggtaaacatt acggcaaaca aacactgaca actgcacaag ttaacgtatc
      901 agcatcagac agctctttga acatcaacgg tgtagaggat tataaatcaa tctttgacgg
      961 tgacggaaaa acgtatcaaa atgtacagca gttcatcgat gaaggcaact acagctcagg
     1021 cgacaaccat acgctgagag atcctcacta cgtagaagat aaaggccaca aatacttagt
     1081 atttgaagca aacactggaa ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa
     1141 agcatactat ggcaaaagca catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag
     1201 cgataaaaaa cgcacggctg agttagcaaa cggcgctctc ggtatgattg agctaaacga
     1261 tgattacaca ctgaaaaaag tgatgaaacc gctgattgca tctaacacag taacagatga
     1321 aattgaacgc gcgaacgtct ttaaaatgaa cggcaaatgg tacctgttca ctgactcccg
     1381 cggatcaaaa atgacgattg acggcattac gtctaacgat atttacatgc ttggttatgt
     1441 ttctaattct ttaactggcc catacaagcc gctgaacaaa actggccttg tgttaaaaat
     1501 ggatcttgat cctaacgatg taacctttac ttactcacac ttcgctgtac ctcaagcgaa
     1561 aggaaacaat gtcgtgatta caagctatat gacaaacaga ggattctacg cagacaaaca
     1621 atcaacgttt gcgccaagct tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa
     1681 agacagcatc cttgaacaag gacaattaac agttaacaaa taagaaaacc ctggcgacta
     1741 gtcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     1801 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaggggtc cccaataatt
     1861 acgatttaaa tttgacataa gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc
     1921 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     1981 ttttcatggc ttgttatgac tgtttttttg tacagcctat gcctcgggca tccaagcagc
     2041 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     2101 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg
     2161 ggcatcattc gcacatgtag gctcggccct gaccaagtca aatccatgcg ggctgctctt
     2221 gatcttttcg gtcgtgagtt cggagacgta gccacctact cccaacatca gccggactcc
     2281 gattacctcg ggaacttgct ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa
     2341 gaagcggttg ttggcgctct cgcggcttac gttctgccca agtttgagca gccgcgtagt
     2401 gagatctata tctatgatct cgcagtctcc ggagagcacc ggaggcaggg cattgccacc
     2461 gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg
     2521 caagcagatt acggtgacga tcccgcagtg gctctctata caaagttggg catacgggaa
     2581 gaagtgatgc actttgatat cgacccaagt accgccacct aacaattcgt tcaagccgag
     2641 atcggcttcc cggccgcgga gttgttcggt aaattggaca acggtccgcg cgttgtcctt
     2701 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     2761 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     2821 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     2881 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     2941 gtaggccggc cgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3001 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3061 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg catccggtaa
     3481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc cgtgaaaggc aggccggtcc
     3661 gtggtggcca cggcctctag gccagatcca gcggcatctg ggttagtcga gcgcgggccg
     3721 cttcccatgt ctcaccaggg cgagcctgtt tcgcgatctc agcatctgaa atcttcccgg
     3781 ccttgcgctt cgctggggcc ttacccaccg ccttggcggg cttcttcggt ccaaaactga
     3841 acaacagatg tgtgaccttg cgcccggtct ttcgctgcgc ccactccacc tgtagcgggc
     3901 tgtgctcgtt gatctgcgtc acggctggat caagcactcg caacttgaag tccttgatcg
     3961 agggataccg gccttccagt tgaaaccact ttcgcagctg gtcaatttct atttcgcgct
     4021 ggccgatgct gtcccattgc atgagcagct cgtaaagcct gatcgcgtgg gtgctgtcca
     4081 tcttggccac gtcagccaag gcgtatttgg tgaactgttt ggtgagttcc gtcaggtacg
     4141 gcagcatgtc tttggtgaac ctgagttcta cacggccctc accctcccgg tagatgattg
     4201 tttgcaccca gccggtaatc atcacactcg gtcttttccc cttgccattg ggctcttggg
     4261 ttaaccggac ttcccgccgt ttcaggcgca gggccgcttc tttgagctgg ttgtaggaag
     4321 attcgatagg gacacccgcc atcgtcgcta tgtcctccgc cgtcactgaa tacatcactt
     4381 catcggtgac aggctcgctc ctcttcacct ggctaataca ggccagaacg atccgctgtt
     4441 cctgaacact gaggcgatac gcggcctcga ccagggcatt gcttttgtaa accattgggg
     4501 gtgaggccac gttcgacatt ccttgtgtat aaggggacac tgtatctgcg tcccacaata
     4561 caacaaatcc gtccctttac aacaacaaat ccgtcccttc ttaacaacaa atccgtccct
     4621 taatggcaac aaatccgtcc ctttttaaac tctacaggcc acggattacg tggcctgtag
     4681 acgtcctaaa aggtttaaaa gggaaaagga agaaaagggt ggaaacgcaa aaaacgcacc
     4741 actacgtggc cccgttgggg ccgcatttgt gcccctgaag gggcggggga ggcgtctggg
     4801 caatccccgt tttaccagtc ccctatcgcc gcctgagagg gcgcaggaag cgagtaatca
     4861 gggtatcgag gcggattcac ccttggcgtc caaccagcgg caccagcggc gcctgagagg
     4921 ggcgcgccca gctgtctagg gcggcggatt tgtcctactc aggagagcgt tcaccgacaa
     4981 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     5041 ttaattaatt gacaccatct cgaggaattc ttgacagcta gctcagtcct aggtataatg
     5101 ctagcaattt ctactgttgt agatagagac cactgcacgg gtctcttttt tttgaggagc
     5161 tcggtacccg gggatcccac tcttgagggc ccaaggcggc cgcgtcgtga ctgg
//
