LOCUS       sequence-204354-        6020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for expression of computationally designed colicin 
            endonuclease and immunity pair, des13.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6020)
  AUTHORS   Netzer R, Listov D, Lipsh R, Dym O, Albeck S, Knop O, Kleanthous C, 
            Fleishman SJ
  TITLE     Ultrahigh specificity in a network of computationally designed 
            protein-interaction pairs.
  JOURNAL   Nat Commun. 2018 Dec 11;9(1):5286. doi: 10.1038/s41467-018-07722-9.
  PUBMED    30538236
REFERENCE   2  (bases 1 to 6020)
  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: "Nat 
            Commun."; date: "2018-12-11"; volume: "9(1):5286. doi"; pages: " 
            10.1038/s41467-018-07722-9"
COMMENT     Sequence Label: pET21d-colEdes13/Imdes13
FEATURES             Location/Qualifiers
     source          1..6020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     501..520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     754..771
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    786..925
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(851..870)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1011..1033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1027..1218)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3070..3089
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3109..3186)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3392..3411
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3495..3514
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3495..3513
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3514..3538
                     /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)."
     RBS             3553..3575
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4247..4264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4317..4335)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4331..4378
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4415..4870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4502..4521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4712..4733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4896..5000
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5001..5069,5070..5861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5219..5238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
       61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      121 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
      181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      721 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      781 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
      841 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      901 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      961 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     1021 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     1081 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     1141 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     1201 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     1261 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     1321 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     1381 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     1441 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     1501 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     1561 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     1621 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     1681 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     1741 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     1801 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     1861 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     1921 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     1981 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     2041 ccagtcggga aactgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2101 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     2161 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     2221 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     2281 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     2341 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     2401 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     2461 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     2521 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     2581 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     2641 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     2701 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     2761 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     2821 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     2881 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     2941 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     3001 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     3061 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     3121 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     3181 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     3241 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     3301 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     3361 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     3421 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     3481 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     3541 tctagaaata attttgttta actttaagaa ggagatatac catggagtct aagcgtaaca
     3601 aaccaggtaa agcgaccggt aaaggtaagc cggtcggtga taaatggctg gacgacgcgg
     3661 gtaaggactc tggtgcgccg attccggacc gcatcgcgga caaactgcgt gacaaagaat
     3721 tcaaaaactt tgacgacttc cgtaaaaagt tctgggaaga agttagcaag gacccggatc
     3781 tgtctaaaca gttcaaacgt tctaaccaga acaatatcaa acagggtatc gcgccgttcg
     3841 cgcgttccaa agaccaggtt ggtggccgtc gtaccttcga actgcaccac gacaagccga
     3901 tctctcagga cggtggtgtt tacgacatga acaacatccg tgttaccacc ccgaaacgcc
     3961 acatcgacat ccaccggggt aagtaaaaat ggaactgaaa cactctatct ctgactacac
     4021 cgaagcggag ttcctcgagt tcgttaagaa aatcatggcg tctaacgacg aaacccagca
     4081 gcgtaaactg gttaccgaat ttgaacgtct gaccgaacac ccggacggtt ctgacctgat
     4141 ctactaccca cgtgacgacc gtgaagactc tccggaaggt atcgttaaag aaatcaagga
     4201 atggcgtgcg gcgaatggta aatctggttt caaacaaggc ctcgagcacc accaccacca
     4261 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     4321 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     4381 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     4441 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4501 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4561 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4621 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4681 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4741 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4801 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4861 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4981 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5041 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5101 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5161 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5221 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5281 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5341 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5401 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5461 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5521 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     5581 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5641 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5701 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5761 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5821 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5881 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5941 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6001 cagaccccgt agaaaagatc
//
