LOCUS       sequence-204351-        6032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for expression of computationally designed colicin 
            endonuclease and immunity pair, des11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  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 6032)
  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-colEdes11/Imdes11
FEATURES             Location/Qualifiers
     source          1..6032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     762..779
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    794..933
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(859..878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1019..1041
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1035..1226)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3078..3097
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3117..3194)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3400..3419
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3503..3522
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3503..3521
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3522..3546
                     /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             3561..3583
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4267..4284
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4337..4355)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4351..4398
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4435..4890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4522..4541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4732..4753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4916..5020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5021..5089,5090..5881)
                     /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(5239..5258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      721 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      781 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
      841 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
      901 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      961 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     1021 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     1081 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     1141 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     1201 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     1261 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     1321 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     1381 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     1441 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     1501 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     1561 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     1621 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     1681 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     1741 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     1801 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     1861 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     1921 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     1981 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     2041 cccgctttcc agtcgggaaa ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2101 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     2161 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     2221 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     2281 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     2341 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     2401 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     2461 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     2521 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     2581 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     2641 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     2701 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     2761 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     2821 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     2881 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     2941 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     3001 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     3061 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     3121 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     3181 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     3241 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     3301 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     3361 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     3421 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     3481 cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
     3541 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca tggagtctaa
     3601 gcgtaacaaa ccgggtaaag cgaccggtaa aggtaagccg gttggtgaca aatggctgga
     3661 cgacgcaggt aaggattctg gtgcgccgat cccggaccgt atcgcggaca agctgcgtga
     3721 caaggaattc aaaaactttg acgacttcaa gaaaaagttc tgggaagaag tttctaaaga
     3781 tccggacctg tctaaacagt tcaaaggttc taaccaggaa tctatcaaaa aaggcctcgc
     3841 gccattcgcg cgtcagaacg accaggtcgg tggtcgtcgt gttttcgaac tgcaccacga
     3901 taaaccgatc tctcaggacg gtggtgttta cgacatgaac aacatccgcg ttaccacccc
     3961 aaaacgtcac atcgacatcc accggggtaa gtaaaaatgg aactgaaaca ctctatctct
     4021 gactacaccg aagcggaatt tctggaattc gttaagaaga tctgcaatct gtctgcgggc
     4081 aacgcgtctg ttgatgaaat ggaaaaagcg gttgacgagt tcgagcgcct gacggagcac
     4141 ccgtctggtt ccgacctcat ctactacccg cgtgacgacc gtgaagactc tccggaaggt
     4201 atcgttaaag aaatcaagga gtggcgtgcg gcgaacggta aatctggttt caaacagggc
     4261 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     4321 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     4381 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     4441 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4501 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4561 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4621 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4681 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4741 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4801 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4861 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4921 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4981 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5041 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5101 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5161 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5221 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5281 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5341 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5401 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5461 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5521 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     5581 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5641 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5701 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5761 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5821 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5881 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6001 gttttcgttc cactgagcgt cagaccccgt ag
//
