LOCUS       sequence-313311-        6758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Cba and Cbi protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6758)
  TITLE     Clone of colicin B and its corresponding immunity protein.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6758)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pKP579
FEATURES             Location/Qualifiers
     source          1..6758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(80..85,86..102,103..108)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2581..2592
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3021..3040)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4893..5552)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5467..5486
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5553..5655)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6047..6064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6181..6726)
                     /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(6261..6280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
       61 taagttgtaa ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat
      121 cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg
      181 tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc
      241 cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc
      301 tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc
      361 gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga
      421 tcatggcgac cacacccgtc ctgtggatcc agaacagtta tgcctgtgcc gttttttgtg
      481 gtatcaggcg agaatagcac gtggtgtgaa tgcctgtttt ttgatcgttt tcacaaaaat
      541 tgaaatccac agtcttgaca gagaaaacac agcggcgtaa tttttgatct gtacataaaa
      601 ccagtggtta tatgtacagt atttaatttt aattgattgt ttttaaagtc aaagaggttt
      661 tctatgagtg ataatgaagg tagtgtaccg acagaaggta ttgattacgg ggacacaatg
      721 gttgtgtggc cgtcaacagg acgaattccg ggcggtgatg tgaaacccgg aggctcatca
      781 ggtctcgctc catccatgcc tccgggatgg ggggattaca gcccacaagg tatcgcactt
      841 gtacaaagtg ttctttttcc tggaattatt cgccggatta ttcttgataa agaacttgaa
      901 gagggagact ggtcgggatg gtctgtcagt gtgcatagcc cctggggaaa cgagaaagtt
      961 tccgctgcac gaacagttct tgaaaatggt ttacgtggtg gtttgccaga accgtctcgc
     1021 ccggctgctg tttcttttgc ccgtctggag cctgcttccg gaaatgagca aaaaattatt
     1081 cgtcttatgg ttacacagca actggagcag gtaacggata tccctgccag ccagttacca
     1141 gcagcgggta ataatgtacc ggtaaaatat cgtctgacgg accttatgca gaatggtaca
     1201 caatatatgg ctattatcgg aggtattccg atgacagtgc cagtagtgga tgccgttcca
     1261 gttccggacc ggagtcgtcc gggaaccaat attaaagatg tttacagtgc ccctgtatca
     1321 ccaaatctac cggacctggt attaagtgtg ggtcagatga atactccagt tcggtctaat
     1381 cccgaaatcc aggaagatgg cgttatttct gagacaggga attatgttga ggctggttat
     1441 acgatgtcca gtaataatca tgatgtcatt gtccgttttc ctgaaggcag tggagtttct
     1501 ccgctatata tttcagccgt ggagattctg gacagtaata gtttaagcca gcgccaggaa
     1561 gccgaaaata acgcaaagga tgacttcaga gtcaagaaag aacaagaaaa tgacgagaag
     1621 acggtcctga caaaaaccag cgaggtcatc attagtgtcg gtgacaaagt cggggaatat
     1681 cttggagata aatacaaggc gctttcccgt gaaattgcag agaatataaa taattttcag
     1741 ggaaaaacga ttcgtagtta tgatgatgca atgtcttcca ttaataagtt aatggctaac
     1801 cccagcctta aaataaatgc aacggacaaa gaagccattg tgaatgcgtg gaaagcattt
     1861 aatgctgagg atatggggaa taaatttgct gcgttgggta aaacgttcaa agcagcagat
     1921 tatgcaataa aggcaaacaa catcagggag aagagtattg agggttacca gactggtaac
     1981 tgggggccat taatgctgga agtcgagtcc tgggttatca gtgggatggc atctgctgta
     2041 gctcttagtt tgttttcttt gacattaggc tcggccctta tagcctttgg tctttcggcc
     2101 acagttgttg gttttgttgg cgtagttatt gcaggtgcta ttggtgcatt tatcgatgat
     2161 aaatttgttg atgagttgaa tcacaagatc ataaaataat taaaaggaag gcagtgttac
     2221 tgcctccctt taaaatattt aatgacaagt aatggcatca gtaaaatcgc atatgtcatg
     2281 gttaaaacag taaagtaaat ggagcaatat aaaataaaca gagtgatgac attatgagat
     2341 aataatttaa aaaaaccacc tgctgttgtt agttccagat tatagaacag tgtgatataa
     2401 atcagaatgg cataaaaaac gggcccccag aaacaggatt gcacaacaag ttttcttgtt
     2461 actggcttta tgatttctct tgtcttatat gtgcaaagaa aaacgagaat acccataact
     2521 ggcgctgtct tgttgtagtt tgacataagt gttgtcatga cgggatcatg caatgatgta
     2581 atagaaggga gaaatgccac cttattatat aagtagtaca gcacttgaga atatggatca
     2641 tttattcgaa gtattaatat agccattaat ggaataatcc cgattatgga aaatgcatat
     2701 aatatttcgt ttgctttctt gttcttatct ttattgctgg tcatatatat acctttttgt
     2761 attttatata tagattatat atcagagttc tcaatatatt tccagtattt aatggagtgt
     2821 acagatatat atcttatata tggtagttca ggagctgccc taccattttt aacggcatat
     2881 caggcaaaca atgttatggg gagcctctct catggggttt gttcgccggc ggaggcgtta
     2941 gccagcggcg aactctggga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     3001 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     3061 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     3121 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     3181 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgacc
     3241 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     3301 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     3361 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
     3421 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
     3481 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta
     3541 cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     3601 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     3661 ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcattgg
     3721 accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg
     3781 gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     3841 ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
     3901 agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
     3961 aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt
     4021 gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     4081 cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     4141 tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     4201 tcgtaaagtc tggaaacgcg gaagtcccct acgtgctgct gaagttgccc gcaacagaga
     4261 gtggaaccaa ccggtgatac cacgatacta tgactgagag tcaacgccat gagcggcctc
     4321 atttcttatt ctgagttaca acagtccgca ccgctgccgg tagctccttc cggtgggcgc
     4381 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     4441 aggtgccggc agcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     4501 aacaagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
     4561 gtggaacacc tacatctgta ttaacgaagc gctaaccgtt tttatcaggc tctgggaggc
     4621 agaataaatg atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc
     4681 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     4741 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     4801 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc
     4861 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     4921 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     4981 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     5041 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     5101 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     5161 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     5221 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     5281 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc
     5341 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     5401 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     5461 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     5521 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     5581 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     5641 cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca
     5701 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
     5761 gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg
     5821 aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt
     5881 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     5941 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     6001 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     6061 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     6121 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     6181 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     6241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     6301 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     6361 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     6421 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6481 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     6541 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     6601 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     6661 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     6721 tctcaagaag atcatcttat taatcagata aaatattt
//
