LOCUS       62056_13210        5087 bp DNA     circular SYN 07-JAN-2021
DEFINITION  Suicide vector pJP5603sacBGm DNA, complete sequence.
ACCESSION   LC599391
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5087)
  AUTHORS   Zoolkefli FIR.M., Moriguchi K, Cho Y, Kiyokawa K, Yamamoto S, Suzuki
            K.
  TITLE     Isolation and Analysis of Donor Chromosomal Genes Whose Deficiency 
            is Responsible for Accelerating Bacterial and Trans-kingdom 
            Conjugations by IncP1 T4SS Machinery
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5087)
  AUTHORS   Zoolkefli FIR.M., Moriguchi K, Cho Y, Kiyokawa K, Yamamoto S, Suzuki
            K.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-DEC-2020) Contact:Kazuki Moriguchi Hiroshima 
            University, Biological Science; 1-3-1 Kagamiyama, Higashi-Hiroshima 
            City, Hiroshima 739-8526, Japan
REFERENCE   3  (bases 1 to 5087)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-DEC-2020) Contact:Kazuki Moriguchi Hiroshima University, 
            Biological Science; 1-3-1 Kagamiyama, Higashi-Hiroshima City, 
            Hiroshima 739-8526, Japan"
FEATURES             Location/Qualifiers
     source          1..5087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        338..366
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             555..1085
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     protein_bind    1417..1438
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1453..1483
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1491..1507
                     /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     1515..1531
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(1544..1600)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1601..1617)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1689..1757
                     /note="beta-galactosidase; LacZ alpha"
     promoter        2049..2494
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2495..3913
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    4053..4237
                     /label=TraJ (RP4 mob)
                     /note="TraJ (RP4 mob)"
     oriT            complement(4270..4379)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      4539..4927
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa
       61 gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
      121 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcata
      181 aaaactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
      241 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
      301 acgcacaccg tggaaacgga tgaaggcacg aacccagttg acataagcct gttcggttcg
      361 taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
      421 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttgtaca
      481 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
      541 atggagcagc aacgatgtta cgcagcagca acgatgttac gcagcagggc agtcgcccta
      601 aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc ggccctgacc
      661 aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcgga gacgtagcca
      721 cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt agtaagacat
      781 tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg gcttacgttc
      841 tgcccaggtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca gtctccggcg
      901 agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat gaggccaacg
      961 cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc gcagtggctc
     1021 tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac ccaagtaccg
     1081 ccacctaaca attcgttcaa gccgagatcg gcttcccggc cgcggagttg ttcggtaaat
     1141 tgtcacaacg ccggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     1201 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     1261 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccccaat
     1321 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1381 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     1441 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1501 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
     1561 ggtcgactct agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac
     1621 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1681 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1741 gcctgaatgg cgaatggggc tcgatcccct cgcgagttgg ttcagctgct gcctgaggct
     1801 ggacgacctc gcggagttct accggcagtg caaatccgtc ggcatccagg aaaccagcag
     1861 cggctatccg cgcatccatg cccccgaact gcaggagtgg ggaggcacga tggccgcttt
     1921 ggtcggtctt gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct
     1981 tgatggagcg catggggacg tgcttggcaa tcacgcgttc gctagaggat cgatcctttt
     2041 taacccatca catatacctg ccgttcacta ttatttagtg aaatgagata ttatgatatt
     2101 ttctgaattg tgattaaaaa ggcaacttta tgcccatgca acagaaacta taaaaaatac
     2161 agagaatgaa aagaaacaga tagatttttt agttctttag gcccgtagtc tgcaaatcct
     2221 tttatgattt tctatcaaac aaaagaggaa aatagaccag ttgcaatcca aacgagagtc
     2281 taatagaatg aggtcgaaaa gtaaatcgcg cgggtttgtt actgataaag caggcaagac
     2341 ctaaaatgtg taaagggcaa agtgtatact ttggcgtcac cccttacata ttttaggtct
     2401 ttttttattg tgcgtaacta acttgccatc ttcaaacagg agggctggaa gaagcagacc
     2461 gctaacacag tacataaaaa aggagacatg aacgatgaac atcaaaaagt ttgcaaaaca
     2521 agcaacagta ttaaccttta ctaccgcact gctggcagga ggcgcaactc aagcgtttgc
     2581 gaaagaaacg aaccaaaagc catataagga aacatacggc atttcccata ttacacgcca
     2641 tgatatgctg caaatccctg aacagcaaaa aaatgaaaaa tatcaagttt ctgaatttga
     2701 ttcgtccaca attaaaaata tctcttctgc aaaaggcctg gacgtttggg acagctggcc
     2761 attacaaaac gctgacggca ctgtcgcaaa ctatcacggc taccacatcg tctttgcatt
     2821 agccggagat cctaaaaatg cggatgacac atcgatttac atgttctatc aaaaagtcgg
     2881 cgaaacttct attgacagct ggaaaaacgc tggccgcgtc tttaaagaca gcgacaaatt
     2941 cgatgcaaat gattctatcc taaaagacca aacacaagaa tggtcaggtt cagccacatt
     3001 tacatctgac ggaaaaatcc gtttattcta cactgatttc tccggtaaac attacggcaa
     3061 acaaacactg acaactgcac aagttaacgt atcagcatca gacagctctt tgaacatcaa
     3121 cggtgtagag gattataaat caatctttga cggtgacgga aaaacgtatc aaaatgtaca
     3181 gcagttcatc gatgaaggca actacagctc aggcgacaac catacgctga gagatcctca
     3241 ctacgtagaa gataaaggcc acaaatactt agtatttgaa gcaaacactg gaactgaaga
     3301 tggctaccaa ggcgaagaat ctttatttaa caaagcatac tatggcaaaa gcacatcatt
     3361 cttccgtcaa gaaagtcaaa aacttctgca aagcgataaa aaacgcacgg ctgagttagc
     3421 aaacggcgct ctcggtatga ttgagctaaa cgatgattac acactgaaaa aagtgatgaa
     3481 accgctgatt gcatctaaca cagtaacaga tgaaattgaa cgcgcgaacg tctttaaaat
     3541 gaacggcaaa tggtacctgt tcactgactc ccgcggatca aaaatgacga ttgacggcat
     3601 tacgtctaac gatatttaca tgcttggtta tgtttctaat tctttaactg gcccatacaa
     3661 gccgctgaac aaaactggcc ttgtgttaaa aatggatctt gatcctaacg atgtaacctt
     3721 tacttactca cacttcgctg tacctcaagc gaaaggaaac aatgtcgtga ttacaagcta
     3781 tatgacaaac agaggattct acgcagacaa acaatcaacg tttgcgccga gcttcctgct
     3841 gaacatcaaa ggcaagaaaa catctgttgt caaagacagc atccttgaac aaggacaatt
     3901 aacagttaac aaataaaaac gcaaaagaaa atgccgatgg gtaccgagcg aaatgaccga
     3961 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4021 gttgggcttc ggaatcgttt tccgggacgc cctcgcggac gtgctcatag tccacgacgc
     4081 ccgtgatttt gtagccctgg ccgacggcca gcaggtaggc cgacaggctc atgccggccg
     4141 ccgccgcctt ttcctcaatc gctcttcgtt cgtctggaag gcagtacacc ttgataggtg
     4201 ggctgccctt cctggttggc ttggtttcat cagccatccg cttgccctca tctgttacgc
     4261 cggcggtagc cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat
     4321 aagggacagt gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg
     4381 gctgacgccg ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat
     4441 atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt
     4501 atgcagcgga aaacggccac gatgcgtccg gcgtagagga tctgaagatc agcagttcaa
     4561 cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     4621 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
     4681 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
     4741 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
     4801 tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac
     4861 gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg
     4921 gctgacatgg gaattagctt cacgctgccg caagcactca gggcgcaagg gctgctaaag
     4981 gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag
     5041 ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagc
//
