LOCUS       sequence-485424-       11131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ColK-based vector; provides a nicking function to permit conjugal 
            transfer and bears a polylinker for insertion of methylase-encoding 
            genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11131)
  AUTHORS   Elhai J, Vepritskiy A, Muro-Pastor AM, Flores E, Wolk CP
  TITLE     Reduction of conjugal transfer efficiency by three restriction 
            activities of Anabaena sp. strain PCC 7120.
  JOURNAL   J Bacteriol. 1997 Mar;179(6):1998-2005.
  PUBMED    9068647
REFERENCE   2  (bases 1 to 11131)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J 
            Bacteriol."; date: "1997-03"; volume: "179(6)"; pages: "1998-2005"
COMMENT     Sequence Label: pRL542
FEATURES             Location/Qualifiers
     source          1..11131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4232..4251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4472..4489
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4730..4752
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ctgcaggtcg actctagagg atccccgggt accgagctcg aattcatcga tatctagatc
       61 tcgagctatc aggctagaga ttttaccgcc aaatcgaacc ttattagagc ggtttaggct
      121 ggaccggcag ttaaaattgg ggcttgagcg gtaaacgagt gagggaattt caggtaagat
      181 acttcggatg aggagcaaaa aggtggttta tacttcctat acccaaaaag gtggtttata
      241 cttcctatac cccggttatc cggtggtttt ctgtttgcgg aataaaacag attagtgaca
      301 gttttctcag aatggcgtgt tccggtgact tctgttttcg tgtgctgcat ccggaagtgt
      361 caggcccgcc ggatatgaaa aaggcctccg gagagtaccg tgactttatt ttcggcacaa
      421 atacaggggt cgatggataa atacggcgat agtttcctga cggatgatcc gtatgtaccg
      481 gcggaagaca agctgcaaac ctgtcagatg gagattgatt taatggcgga tgtgctgaga
      541 gcaccgcccc gtgaatccgc agaactgatc cgctatgtgt ttgcggatga ttggccggaa
      601 taaataaagc cgggcttaat acagattaag cccgtatagg gtattattac tgaataccaa
      661 acagcttacg gaggacggaa tgttacccat tgagacaacc agactgcctt ctgattatta
      721 atatttttca ctattaatca gaaggaataa ccatgaattt tacccggatt gacctgaata
      781 cctggaatcg cagggaacac tttgcccttt atcgtcagca gattaaatgc ggattcagcc
      841 tgaccaccaa actcgatatt accgctttgc gtaccgcact ggcggagaca ggttataagt
      901 tttatccgct gatgatttac ctgatctccc gggctgttaa tcagtttccg gagttccgga
      961 tggcactgaa agacaatgaa cttatttact gggaccagtc agacccggtc tttactgtct
     1021 ttcataaaga aaccgaaaca ttctctgcac tgtcctgccg ttattttccg gatctcagtg
     1081 agtttatggc aggttataat gcggtaacgg cagaatatca gcatgatacc agattgtttc
     1141 cgcagggaaa tttaccggag aatcacctga atatatcatc attaccgtgg gtgagttttg
     1201 acggatttaa cctgaacatc accggaaatg atgattattt tgccccggtt tttacgatgg
     1261 caaagtttca gcaggaaggt gaccgcgtat tattacctgt ttctgtacag gttcatcatg
     1321 cagtctgtga tggctttcat gcagcacggt ttattaatac acttcagctg atgtgtgata
     1381 acatactgaa ataaattaat taattctgta tttaagccac cgtatccggc aggaatggtg
     1441 gctttttttt atattttaac cgtaatctgt aatttcgttt cagactggtt caggatcact
     1501 gtacgataat gcccccgcag tttggtaata cccttaataa aaaagaaaca gcaaagactg
     1561 acagcaataa taataaagta agcagtaaca ataatattaa caacaccaga tgcagttata
     1621 ataatagtat ttaagacacc agaaagactg ctgcgacagt cattttgaac aacaccaaaa
     1681 tgccgtaaag gcagtagtaa caacaccagt gaaaacatca cgatagcata gtgatatgcc
     1741 tgagtgtgtg taattaaaca ataaataaac cgccatatat aacagaagat agtattctga
     1801 atggcatgct tttctgttca gtataaacat atcatcccgg ttggtataag gatgatatat
     1861 aataagttaa gctgaacaca tatttatttt ggttttattt tacaaataaa gtaagacgat
     1921 ccgttaagtc aaagcggggt atatttatta taccctgctt ttttatttgt ccgccgggcg
     1981 cggataatgg atcagattat gcagtgtcac aatggcctta ccgggattgg cgtaagcgtg
     2041 cgggatatcc gcatggaagc gcagggattc cccggcagaa acggtgtgcc actcatcccc
     2101 cagccgcagt tgtaatgcgc cttccagtac aatgacatgt tctctggttc tgaaatccat
     2161 ccctgtcggt gttgcttatg cagtctggtc gggactcggc gtcgtcataa ttacagccat
     2221 tgcctggttg cttcatgggc aaaagcttga tgcgtggggc tttgtaggta tggggctcat
     2281 aattgctgcc tttttgctcg cccgatcccc atcgtggaag tcgctgcgga ggccgacgcc
     2341 atggtgacgg tgttcggcat tctgaatctc accgaggact ccttcttcga tgagagccgg
     2401 cggctagacc ccgccggcgc tgtcaccgcg gcgatcgaaa tgctgcgagt cggatcagac
     2461 gtcgtggatg tcggaccggc cgccagccat ccggacgcga ggcctgtatc gccggccgat
     2521 gagatcagac gtattgcgcc gctcttagac gccctgtccg atcagatgca ccgtgtttca
     2581 atcgacagct tccaaccgga aacccagcgc tatgcgctca agcgcggcgt gggctacctg
     2641 aacgatatcc aaggatttcc tgaccctgcg ctctatcccg atattgctga ggcggactgc
     2701 agatggaatg cgctttgtta cgccagtcag aacaatcaat gccggaccga accgcaaata
     2761 ctttggtaat aacagaggga tcacctggta caactttgtg tccgatcagt attccggctt
     2821 tcatggcatc gttataccgg ggacgctgag ggactctatc tttgtgctgg aaggccttct
     2881 ggaacaggag accgggctga atccaaccga aattatgacc gatacagcag gtaccagcga
     2941 acttgtcttt ggccttttct ggctgctggg ataccagttt tctccacgcc tggctgatgc
     3001 cggtgcttcg gttttctggc gaatgggcca tgatgccaac tatggcgtgc tgaatgatat
     3061 tgccagaggg caatcagatc cccgaaaaat agtccttcag tgggacgaaa tgatccggac
     3121 cgctggctcc ctgaaactgg gcaaagtaca ggcttcagtg ctggtccgtt cattgctgaa
     3181 aagtgaacgt ccttccggac tgactcaggc aatcattgaa gtggggcgca tcaacaaaac
     3241 gctgtatctg cttaattata ttgatgatga agattaccgc cggcgcattc tgacccagct
     3301 taatcgggga gaaagccgcc atgccgttgc cagagccatc tgtcacggtc aaaaaggtga
     3361 gataagaaaa cgatataccg acggtcagga agatcaactg ggcgcactgg ggctggtcac
     3421 taacgccgtc gtgttatgga acactatgta tatgcaggca gccctggatc atctccgggc
     3481 gcaaggtgaa acactgaatg atgaagatat cgcacgcctc tccccgcttt gccacggaca
     3541 tatcaatatg ctcggccatt attccttcac gctggcagaa ctggtgacca aaggacatct
     3601 gagaccatta aaagaggcgt cagaggtaga aaacgttgct taacgtgagt tttcgttcca
     3661 ctgagcgtca gacccctgta caaggaggaa ccgttgcacc atcttcgtct tctgaactga
     3721 cggggatcgc ggttcagtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3781 cgtaatctgc tgcctgtaaa cgaaaaaacc gcctggggag gcggtttagt cgaaggttaa
     3841 gtcagttggg gactgcttaa ccgggtaact ggctttagtg gagcacagat accaaatgct
     3901 gtccttccag tgtagccgct ataaggcttc aacttcaaga gttctgcgta tctaaatcca
     3961 ctatccagtt accaatggct gctgccagtg gcgttaagtc gtgtcttacc gggttggact
     4021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4081 agcccagctt ggagcgaacg acctacaccg aactgagata ccaacagcgt gagctatgag
     4141 aaagcgccac gcttcccaaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4201 gaacaggaga gcgcacgagg gagccaccag ggggaaacgc ctggtatctt taagtcctgt
     4261 cgggtttcgc cacctctgac ttgagcgtcg gtttttgtga tgctcgtcag gggggcggag
     4321 cctatggaaa aacgccagct gcgtggcctt ttccgggttc ttgttttttg ctcacatgtt
     4381 tcttttggtg ttatcccctg attctgtgga taaccgcatt accgtctttg agtgagctga
     4441 taccgctcgc cgcagccgaa cgaccgagcg gagcgagtca gtgagcgagg aagcggaaaa
     4501 gcgcctggac gagcattttc tccttacgca tctgtgcggc atttcacacc cggcatggtg
     4561 cacttttcat acaatcctca ctgatgccaa atgattaagc cagtatacac tccgctatcg
     4621 ctacgtgact gggtcatggc tgcgccccga cacccgctaa aacccgctga cgcgccctga
     4681 cgggcttgtc tgctcccggc atccgctcac aggccagctg tgaccgtctc cgggagctgc
     4741 atgtgtcaga ggttttcacc gttatccccg aagcgtgcga ggcagctgcg gtaaagctcg
     4801 tcagcgtgat cgtgaagcgc ttcgcagatg tcagcctggt cgtcggcgtc cagttcattc
     4861 agtttttcca gcagggtcag cgtctggctt ctgatgaagc gggccatgtt aagggcggtt
     4921 ttttcctgtt tagtcacagt tctgtttctc catgtgtggg gagtctgtcg gaggttttta
     4981 atgataccgg tgaaacgcga aaggatgctc acgatacggg tcacggatga ggaacatgcg
     5041 cggctgctgg cacgttgtga gggaaaacaa cttgcgtcat ggatgcgtaa ggtctgcctg
     5101 ggggcaccgc catcgaaaac gtcaggtctt ccgacactgg caccgccgct gttgcgccag
     5161 ttcgcttcgg tcggcaataa cctgaatcag atagcccgga agataaacag cggacagtgg
     5221 tcggggcatg accgggttca tgtggtggct gcactgatgg ccatcggacg tgaactgagt
     5281 gagctgaggg atgaggtccg taaacaggga gaacgggatg atagttaagt ttcatgccag
     5341 gggaaaaggc ggtggcagtg gtccggttga ttacctgctg ggccgggagc gtaaccggga
     5401 aggggcgaga gttctgcggg gtgctcccga agaggtgcgg gaactcatcg atgccacgcc
     5461 gtttgcgaaa aaatacacct cgggggttct gtcttttgca gagcagaccc tgccaccggg
     5521 agagcgtgaa agggtgatgg agagctttga gcgggtactg atgcccggtc ttgaaaaaaa
     5581 tcagtacagc atcctgtggg tggagcacca ggacaagggg cgtcttgagc tgaattttgt
     5641 catcccgaat atggagctgg cgagcggaaa acggctccag ccctattacg accgagcgga
     5701 ccgtccgcga atcaatgcct ggcagacact ggtgaatcac cattacgggc tgcacgaccc
     5761 gaacgcaccg gagaacaggc ggacacttgt caccccgaat aatctgccga aagcgaaaca
     5821 ggaggccgca gaagcgatta cgaggggact ggaggctctt taccatgccg gagcgctaaa
     5881 aacgcgtcag gatgtcactg aggcgcttac agcggcaggt tttgaggttg tgaggacaac
     5941 caggagcagt atcagtatcg ccgacccgga gggtgggcgt aatctcaggc tgaaaggggc
     6001 gctgtatgag caatctttta caaatgggga cagacttcga gaagaaactg aaagagcgtg
     6061 ccgtctctac agagaacatg ctgaacagcg agttcaggaa gctcgaagcg tctgtagaca
     6121 gggaactgaa cttaaacggg cagaaaatca gagacgctat cagcgcacac acagcagcgg
     6181 tgaaagaaca actggagacg ctgaacagtg ccgtgaacgc gcagttcagc gccacagaga
     6241 acgaactggc ccggcggcag gaagagctgc tctggaagct ggtcaaaggg cggatactgt
     6301 acccgtcgct gacggcgctt tgcgtcacgg gcgggatttt tctggcgtca tgggggctga
     6361 ttcagtggca ggaaagccgg attgcggcga acatcctggc tatcagggac caggaagaga
     6421 cgctggcgaa actccgggaa aaaacatggg gtgtgacgta tcacgagggc aggaacggga
     6481 aatttctagt gttgccgtcc ggggtgaagg gggaaaacaa ctggacggtg gagaagaaga
     6541 acgcagtaag actggtcagg gagtaagtga gtatgacaga gctggaaatg cacttactga
     6601 acgccttaga gcaattgcag caggactata tgcagcggct gaaagactgg gagagcgcct
     6661 tcacggattt gcagcatatg ttcgagctga cacgacagga gaacgggcaa ctgagggaac
     6721 agtgcgacga cttgagtcag caggtgcaga gattagccgg gcaggtggac cacttgagtc
     6781 ggttatttat cacgaacagc agataaggga tgaacaggaa aggcaggcac gcctggaacg
     6841 ggagcgtgcg cggtcctggc atgaagagca ggaacgggaa gcacgggtat ggcgtggccc
     6901 gtcaataggt atgtgacggt gagttgttcc gggaggttgc cagatggtac cggaacgcgt
     6961 taaaaacgca tccgggcgcg attctggcgt gtcgtttttg ttgctgtgtt ttgcctcttt
     7021 gccagatcgc gctcattcct cagcctcatc ccgcgatgtc tgcgtatgtc atcaactgaa
     7081 aagatggaaa aaaaacagtg tcatgaggtt tacgcaggca aaaccagcgt tattcacatg
     7141 gctgaatagc tatgcatacg gttaaaattt atcaggtgcg atcgcggcag cttttcgggt
     7201 ggtttgttgc catttttacc tgtcagcagc cgtgatcgcg ctgaacgcat ttcagcggtg
     7261 cgtacaatta agggattatg gtaaatccct tactgtctgc cctcgtatcc atcgagataa
     7321 accgcagaaa tggtgtctgc cattagtttc atttttttcg gcgaaggcat atcgctgttt
     7381 gtgtattttc tgccagcaaa ggctgttgtg ctccggagtg agtgaaaaaa tttttctgag
     7441 tggaattctt ttacacgttg ctgtgccgtt tctgctgata ctccggctgc tttgtctttc
     7501 atccccatat cccggaactg ctgaaagaac gggagcatca tacgtatggt gtctgctttc
     7561 tgggtgtaga agttaggatc aaatccatca tctgaagcat aaacataatg ccgggtatgt
     7621 cgttccaggc tggtctttgg tgtgcatcct gtaagagcta ttgttgctgt tatcagaggt
     7681 atcagaagca gcctggcagc ggcagatctc ataaacatct ccttgcgttt ttaatcttga
     7741 tactcaggga gttatatctc cgaagagtat ctcttatcat acgtcggttt actcccaatg
     7801 tgatttgatg gtcaatgagt caggaataat cccgtgaaaa aatgtgtgcc agtcctgctt
     7861 tttagtgttc tttctgcttc ggttgccagt catgcggaca gcaatgtttt tcaacaaaag
     7921 gccgagaaga caaaaaacgg cattgttgcg gatgttaatg ccagacagaa aatgttcagt
     7981 aatagactga agatacaact tcagatcgat gagttgagag cagagctggc tggcaccaga
     8041 gatgcccaaa aaagagagga tttgcgtaaa aaactggagt ctctggagcg gcaaaagaaa
     8101 tccttacgct gatgggggcg tattttcccc ggtttcttcc ataacctttt taccgttcag
     8161 atatttacgg gcgaggcgaa tgcgtcttgc ctggtatacc gccgagtaag cctgggtggg
     8221 attcacacca ttactgattt cactgaccac atgacgctga tcagccagtg tcagcgtgta
     8281 aacctgtttc tggatgaggt cacgggcttt gtcatgctca tatccggcag caacaagggt
     8341 gtcgagaagt cgtttgtctg ttttgttcag tttatggcgg ggtgatttgc tcacatcgtt
     8401 ctccgttttt tcatgatcca acagttttat atctcatcct gatttgctcc ggtgttctgt
     8461 ttcgacgttc tcaatgcctg cctccccata gtgtgatcct tattgcaaaa atgaaaatcc
     8521 atgctcttga catggacaat gctgagtagt aggtttttac tgtacataaa accagtggtt
     8581 atatgtacag tatattgttt ttaatttatt gttttaatta tcaaagagga atttatggct
     8641 aaagaactaa gtggatatgg accaactgct ggtgagtcta tggggggaac aggggcgaat
     8701 ctgaatcagc aaggtggaaa taataatagt aatagtgggg ttcactgggg tggtggctct
     8761 gggcatggta ataatggtgg tcagggaaat tcaaattcat ctggttctac atcaactgtg
     8821 atgaaaaccg gtgagtcata tctgactccg tggggtgatg ttgtcattaa taatgatggc
     8881 cttccggtga tgaatggtat tgttatgaca gaagagaatt caactctggt tgataatcca
     8941 tttggagggg tctccagagt tcttaattct ttaattagtg atatgccatc actttttgca
     9001 gaaagctcag gaaataacaa caataacact gcttcagtta atactgcacc aacgaatgct
     9061 caggtcagtg atatggataa gagttcgaag gtggtgagta atgtaattaa tgaaaaacaa
     9121 aagcagaaga ataaaattgc tacacagatt agtgaaaaac agaagaaaat cgaggagatg
     9181 aaaaaagtat tcaagcatca ttcttatcat ggtatcactg atcttgagag ggatgttgat
     9241 gagttgcaaa aaaaatcaaa tcagcttgat gcagatatta gcaaacttaa ttcttataaa
     9301 aatacattgc aaagtaaaat tggtgatgtg aataaacaga aggaagctga agaaaaggct
     9361 cgtgaaaatg cagaggtcgc tgaacatgaa actcttaatg aagagaaaca ggcggtggcg
     9421 gaagcagaga agcgtttagc tgaggcgaaa gctgaactgg cgaaggctga aagtgatgta
     9481 cagagtaagc aagctactgt ttccagagtt gcaggagagc ttgaaaacgc tcagaagagt
     9541 gttgacgtga aggtcaccgg atttccggga tggcgtgacg ttcagaaaaa actacagcgt
     9601 cagcttgaag ctaagcaggc tgagtactcc gcagttgaaa atgaacttaa gaatgctgtt
     9661 agtttcaggg atggtaaggc cgctgaggtt aaagaggcag agcagaaatt aaaagaagct
     9721 caggatgctc ttgaaaagag ccagataaaa gatgctgttg atactatggt tggattttat
     9781 caatatataa ccgaacaata tggagaaaaa tacgcaaaaa ttgctcagga tttagcggaa
     9841 aagtctaaag gaaagaaaat tcaaggtgtt gatgaagctc tggcagcatt tgaaaagtat
     9901 aagaatgtac tggataagaa attcagtaag gttgatcggg atgctatttt taatgcgtta
     9961 gagtctgtta attatgatga gttatctaag aatttaacga agatatcaaa gtcgcttaaa
    10021 atcaccagtc gagtatcttt tctatatgat gttggttctg attttaaaaa tgcaatagaa
    10081 actggtaact ggcgaccttt gtttgtcacg ctggagaaaa gtgcggtcga tgtaggtgtg
    10141 gcaaaaattg tggcattaat gtttagtttt attgttggtg tgcctcttgg tttctggggg
    10201 attgcaattg tcacaggtat tgtttcttct tacatagggg atgatgagtt gagcaaactt
    10261 aatgagttgt tagggattta atttctcttt agagtgtaat atatgaaact tggcaatcct
    10321 actgggattg ccattaataa ataaaacaat gcagataact tgcataagta ccagggttct
    10381 ttttttaact ttgtattttc tttgatatat ctagatatgg ttaatttaga gtatggatat
    10441 agcaatacat gtattgatga tataaatata aataacaaag gagtgttatc attgtcgtta
    10501 aatattaaaa taagaatcca tggtataagt gattcaggta aattatgtag gtagtatttt
    10561 aaatgcataa gtatttttac ttttgttgga aagaaatatg aaaataataa tttttctaat
    10621 tacaatatta ttcttgtcag gatgtcaagc aaactatatt cgtgatgtac ggggtctgac
    10681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    10741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    10801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    10861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    10921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag atccacgctc accggctcca
    10981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    11041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    11101 gttaatagtt tgcgcaacgt tgttgccatt g
//
