LOCUS       sequence-204343-        6029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for expression of computationally designed colicin 
            endonuclease and immunity pair, des3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6029)
  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 6029)
  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-colEdes3/Imdes3
FEATURES             Location/Qualifiers
     source          1..6029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     756..773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    788..927
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(853..872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1013..1035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1029..1220)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3072..3091
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3111..3188)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3394..3413
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3497..3516
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3497..3515
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3516..3540
                     /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             3555..3577
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4258..4275
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4328..4346)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4342..4389
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4426..4881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4513..4532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4723..4744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4907..5011
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5012..5080,5081..5872)
                     /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(5230..5249)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      781 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
      841 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
      901 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
      961 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     1021 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     1081 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     1141 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     1201 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     1261 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     1321 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     1381 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     1441 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     1501 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     1561 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     1621 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     1681 acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     1741 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     1801 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     1861 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     1921 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     1981 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     2041 ttccagtcgg gaaactgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2101 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     2161 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     2221 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     2281 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     2341 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     2401 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     2461 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     2521 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     2581 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     2641 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     2701 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     2761 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     2821 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     2881 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     2941 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     3001 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     3061 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     3121 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     3181 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     3241 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     3301 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     3361 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     3421 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     3481 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     3541 cctctagaaa taattttgtt taactttaag aaggagatat accatggaga gtaaacggaa
     3601 taagccaggg aaggcgacag gtaaaggtaa accagttggt gataaatggc tggacgacgc
     3661 gggtaaagac tctggtgctc cgatcccaga tcgtattgcg gacaaactgc gtgacaaaga
     3721 attcaaaaac ttcgacgact tccgtaagaa attctggaaa gaagttgcga aagacccgga
     3781 tctggcgaag cagttctcta aagcgaacca gcgcaacatc aaagatggta acgcgccgtt
     3841 tgcgcgtgag tctgaccagg tgggtggtcg caccacctac gaactgcacc acgacaaacc
     3901 gatctctcag gacggtggtg tttacgacat gaacaacatc agagtgacca cacctaagcg
     3961 acatattgat attcatcggg gtaagtaaaa atggaactga aacatagtat tagtgattat
     4021 accgaggctg aatttctgga gtttgtgaaa gacatcttcc gtctgtctcg tccgcaggac
     4081 aacgacctgc agatcaaact ggttctggaa ttcaaacgcc tgactgaaca cccggacggt
     4141 tccgacctga tctactaccc gcgttctgac cgcgaagact ctccggaagg tatcgttaaa
     4201 gaaatcaaag aatggcgagc tgctaacggt aagtcaggat ttaaacaggg cctcgagcac
     4261 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     4321 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     4381 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
     4441 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4501 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4561 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4621 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4681 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4741 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4801 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4861 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4921 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4981 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5041 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5101 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5161 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5221 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5281 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5341 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5401 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5461 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5521 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     5581 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5641 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5701 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5761 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5821 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5881 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5941 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6001 ccactgagcg tcagaccccg tagaaaaga
//
