LOCUS       sequence-337304-        6776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express colicinB-His6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6776)
  TITLE     Postle Lab Plasmids
REFERENCE   2  (bases 1 to 6776)
  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"
COMMENT     Sequence Label: pKP864
FEATURES             Location/Qualifiers
     source          1..6776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1534..1551
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1936..1947
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2376..2395)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4248..4907)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4822..4841
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4908..5010)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5402..5419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5536..6081)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5616..5635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6193..6198,6199..6215,6216..6221)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 atgagtgata atgaaggtag tgtaccgaca gaaggtattg attacgggga cacaatggtt
       61 gtgtggccgt caacaggacg aattccgggc ggtgatgtga aacccggagg ctcatcaggt
      121 ctcgctccat ccatgcctcc gggatggggg gattacagcc cacaaggtat cgcacttgta
      181 caaagtgttc tttttcctgg aattattcgc cggattattc ttgataaaga acttgaagag
      241 ggagactggt cgggatggtc tgtcagtgtg catagcccct ggggaaacga gaaagtttcc
      301 gctgcacgaa cagttcttga aaatggttta cgtggtggtt tgccagaacc gtctcgcccg
      361 gctgctgttt cttttgcccg tctggagcct gcttccggaa atgagcaaaa aattattcgt
      421 cttatggtta cacagcaact ggagcaggta acggatatcc ctgccagcca gttaccagca
      481 gcgggtaata atgtaccggt aaaatatcgt ctgacggacc ttatgcagaa tggtacacaa
      541 tatatggcta ttatcggagg tattccgatg acagtgccag tagtggatgc cgttccagtt
      601 ccggaccgga gtcgtccggg aaccaatatt aaagatgttt acagtgcccc tgtatcacca
      661 aatctaccgg acctggtatt aagtgtgggt cagatgaata ctccagttcg gtctaatccc
      721 gaaatccagg aagatggcgt tatttctgag acagggaatt atgttgaggc tggttatacg
      781 atgtccagta ataatcatga tgtcattgtc cgttttcctg aaggcagtgg agtttctccg
      841 ctatatattt cagccgtgga gattctggac agtaatagtt taagccagcg ccaggaagcc
      901 gaaaataacg caaaggatga cttcagagtc aagaaagaac aagaaaatga cgagaagacg
      961 gtcctgacaa aaaccagcga ggtcatcatt agtgtcggtg acaaagtcgg ggaatatctt
     1021 ggagataaat acaaggcgct ttcccgtgaa attgcagaga atataaataa ttttcaggga
     1081 aaaacgattc gtagttatga tgatgcaatg tcttccatta ataagttaat ggctaacccc
     1141 agccttaaaa taaatgcaac ggacaaagaa gccattgtga atgcgtggaa agcatttaat
     1201 gctgaggata tggggaataa atttgctgcg ttgggtaaaa cgttcaaagc agcagattat
     1261 gcaataaagg caaacaacat cagggagaag agtattgagg gttaccagac tggtaactgg
     1321 gggccattaa tgctggaagt cgagtcctgg gttatcagtg ggatggcatc tgctgtagct
     1381 cttagtttgt tttctttgac attaggctcg gcccttatag cctttggtct ttcggccaca
     1441 gttgttggtt ttgttggcgt agttattgca ggtgctattg gtgcatttat cgatgataaa
     1501 tttgttgatg agttgaatca caagattata aaacaccatc accatcacca ttaattaaaa
     1561 ggaaggcagt gttactgcct ccctttaaaa tatttaatga caagtaatgg catcagtaaa
     1621 atcgcatatg tcatggttaa aacagtaaag taaatggagc aatataaaat aaacagagtg
     1681 atgacattat gagataataa tttaaaaaaa ccacctgctg ttgttagttc cagattatag
     1741 aacagtgtga tataaatcag aatggcataa aaaacgggcc cccagaaaca ggattgcaca
     1801 acaagttttc ttgttactgg ctttatgatt tctcttgtct tatatgtgca aagaaaaacg
     1861 agaataccca taactggcgc tgtcttgttg tagtttgaca taagtgttgt catgacggga
     1921 tcatgcaatg atgtaataga agggagaaat gccaccttat tatataagta gtacagcact
     1981 tgagaatatg gatcatttat tcgaagtatt aatatagcca ttaatggaat aatcccgatt
     2041 atggaaaatg catataatat ttcgtttgct ttcttgttct tatctttatt gctggtcata
     2101 tatatacctt tttgtatttt atatatagat tatatatcag agttctcaat atatttccag
     2161 tatttaatgg agtgtacaga tatatatctt atatatggta gttcaggagc tgccctacca
     2221 tttttaacgg catatcaggc aaacaatgtt atggggagcc tctctcatgg ggtttgttcg
     2281 ccggcggagg cgttagccag cggcgaactc tgggatcctc tacgccggac gcatcgtggc
     2341 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     2401 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
     2461 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc
     2521 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa
     2581 gggagagcgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     2641 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     2701 acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac
     2761 gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt
     2821 cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc
     2881 cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat
     2941 tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag
     3001 gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc ttaccagcct
     3061 aacttcgatc attggaccgc tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg
     3121 gaacgggttg gcatggattg taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg
     3181 tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac
     3241 ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa
     3301 accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg cacgcggcgc
     3361 atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     3421 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     3481 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     3541 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cccctacgtg ctgctgaagt
     3601 tgcccgcaac agagagtgga accaaccggt gataccacga tactatgact gagagtcaac
     3661 gccatgagcg gcctcatttc ttattctgag ttacaacagt ccgcaccgct gccggtagct
     3721 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     3781 tgcaactcgt aggacaggtg ccggcagcgc ccaacagtcc cccggccacg gggcctgcca
     3841 ccatacccac gccgaaacaa gcgccctgca ccattatgtt ccggatctgc atcgcaggat
     3901 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctaa ccgtttttat
     3961 caggctctgg gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc
     4021 ctgagcaaac tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     4081 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     4141 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     4201 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     4261 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     4321 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     4381 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     4441 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     4501 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     4561 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     4621 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     4681 gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     4741 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     4801 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     4861 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     4921 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     4981 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag
     5041 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
     5101 acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt
     5161 atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc
     5221 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag
     5281 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     5341 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     5401 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     5461 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     5521 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     5581 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5641 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     5701 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     5761 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     5821 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     5881 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     5941 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     6001 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     6061 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
     6121 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     6181 taattctcat gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt
     6241 aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct
     6301 cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac tgccgggcct
     6361 cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct
     6421 atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg
     6481 ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc
     6541 gaccacaccc gtcctgtgga tccagaacag ttatgcctgt gccgtttttt gtggtatcag
     6601 gcgagaatag cacgtggtgt gaatgcctgt tttttgatcg ttttcacaaa aattgaaatc
     6661 cacagtcttg acagagaaaa cacagcggcg taatttttga tctgtacata aaaccagtgg
     6721 ttatatgtac agtatttaat tttaattgat tgtttttaaa gtcaaagagg ttttct
//
