LOCUS       sequence-200638-        4815 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses colicin E1 and E1 immunity protein without E1 lysis (kil) 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4815)
  AUTHORS   Kienker PK, Jakes KS, Finkelstein A
  TITLE     Identification of channel-lining amino acid residues in the 
            hydrophobic segment of colicin Ia.
  JOURNAL   J Gen Physiol. 2008 Dec;132(6):693-707. doi: 10.1085/jgp.200810042.
  PUBMED    19029376
REFERENCE   2  (bases 1 to 4815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1085/jgp"; 
            journalName: "J Gen Physiol"; date: "2008-12"; volume: "132"; issue:
            "6"; pages: "693-707"
COMMENT     Sequence Label: pKSJ331
FEATURES             Location/Qualifiers
     source          1..4815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2056..2072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2056..2072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2069..2091)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2080..2096
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(join(2104..2110,2111..2128,2129..2134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2149..2170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2287..2304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2458..3046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2538..2557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3217..4008,4009..4077))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3840..3859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4078..4182)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4250..4268
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4306..4328)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4428..4447
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4641..4663
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4655..4672
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4656..4672
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atggaaaccg cggtagcgta ctataaagat ggtgttcctt atgatgataa gggacaggta
       61 attattactc ttttgaatgg tactcctgac gggagtggct ctggcggcgg aggtggaaaa
      121 ggaggcagta aaagtgaaag ttctgcagct attcatgcaa ctgctaaatg gtctactgct
      181 caattaaaga aaacacaggc agagcaggct gcccgggcaa aagctgcagc ggaagcacag
      241 gcgaaagcaa aggcaaacag ggatgcgctg actcagcgcc tgaaggatat cgtgaatgag
      301 gctcttcgtc acaatgcctc acgtacgcct tcagcaacag agcttgctca tgctaataat
      361 gcagctatgc aggcggaagc agagcgtttg cgccttgcga aagcagaaga aaaagcccgt
      421 aaagaagcgg aagcagcaga aaaggctttt caggaagcag aacaacgacg taaagagatt
      481 gaacgggaga aggctgaaac agaacgccag ttgaaactgg ctgaagctga agagaaacga
      541 ctggctgcat tgagtgaaga agctaaagct gttgagatcg cccaaaaaaa actttctgct
      601 gcacaatctg aagtggtgaa aatggatgga gagattaaga ctctcaattc tcgtttaagc
      661 tccagtatcc atgcccgtga tgcagaaatg aaaacgctcg ctggaaaacg aaatgaactg
      721 gctcaggcat ccgctaaata taaagaactg gatgagctgg tcaaaaaact atcaccaaga
      781 gccaatgatc cgcttcagaa ccgtcctttt tttgaagcaa ccagacgacg ggttggggcc
      841 ggtaagatta gagaagaaaa acaaaaacag gtaacagcat cagaaacacg tattaaccgg
      901 ataaatgctg atataactca gatccagaag gctatttctc aggtcagtaa taatcgtaat
      961 gccggtatcg ctcgtgttca tgaagctgaa gaaaatttga aaaaagcaca gaataatctc
     1021 cttaattcac agattaagga tgctgttgat gcaacagtta gcttttatca aacgctgact
     1081 gaaaaatatg gtgaaaaata ttcgaaaatg gcacaggaac ttgctgataa gtctaaaggt
     1141 aagaaaatcg gcaatgtgaa tgaagctctc gctgcttttg aaaaatacaa ggatgtttta
     1201 aataagaaat tcagcaaagc cgatcgtgat gctattttta atgcgttggc atcggtgaag
     1261 tatgatgact gggctaaaca tttagatcag tttgccaagt acttgaagat tacggggcat
     1321 gtttcttttg gatatgatgt ggtatctgat atcctaaaaa ttaaggatac aggtgactgg
     1381 aagccactat ttcttacatt agagaagaaa gctgcagatg caggggtgag ttatgttgtt
     1441 gctttacttt ttagcttgct tgctggaact acattaggta tttggggtat tgctattgtt
     1501 acaggaattc tatgctccta tattgataag aataaactta atactataaa tgaggtgtta
     1561 gggatttaat tattctttat tgatataaaa agtcctagca atccaaatgg gattgctagg
     1621 accaaacaaa gtagattata tagcataaat aggtttaatt ttgctacggg ggcgttattt
     1681 aggttttttc ttctttcgaa aaaatctttc tttatgaagt taaaagctat gtattcaata
     1741 gcatattttg aatatggaca tagaatagtg cttatcacta ttgcatatag catcttatct
     1801 gacacaagga aataataccc ttcgctgttt tttgttataa ggtatatata tataagtgtg
     1861 cagtacaggc caaataaaat attttttatg tagtatctta agctcataaa ttaaacctcg
     1921 ccatatattc ttttcatttt ataaggatcg agttatgagg aaaagatttt ttgtgggaat
     1981 attcgcgata aacctccttg ttggatggga tcctctagag tcgacctgca ggcatgcaag
     2041 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2101 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2161 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2221 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2281 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2341 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2401 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2461 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2521 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2581 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2641 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2701 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2761 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2821 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2881 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2941 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3001 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3061 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3121 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3181 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3241 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3301 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3361 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3421 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3481 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3541 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3601 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3661 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3721 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3781 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3841 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3901 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3961 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4021 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4081 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4141 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4201 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4261 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4321 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4381 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4441 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     4501 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     4561 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     4621 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcga
     4681 gctcggtacc ggaagtccac agtcttgaca gggaaaatgc agcggcgtag cttttatgct
     4741 gtatataaaa ccagtggtta tatgtacagt atttattttt aacttattgt tttaaaagtc
     4801 aaagaggatt ttata
//
