LOCUS       sequence-204342-        6020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for expression of computationally designed colicin 
            endonuclease and immunity pair, des2.
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-colEdes2/Imdes2
FEATURES             Location/Qualifiers
     source          1..6020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     761..778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    793..932
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(858..877)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1018..1040
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1034..1225)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3077..3096
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3116..3193)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3399..3418
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3502..3521
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3502..3520
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3521..3545
                     /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             3560..3582
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4254..4271
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4324..4342)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4338..4385
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4422..4877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4509..4528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4719..4740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4903..5007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5008..5076,5077..5868)
                     /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(5226..5245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      781 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      841 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
      901 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
      961 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     1021 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     1081 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     1141 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     1201 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     1261 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     1321 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     1381 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     1441 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     1501 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     1561 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     1621 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     1681 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     1741 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     1801 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     1861 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     1921 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     1981 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     2041 ccgctttcca gtcgggaaac tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2101 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     2161 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     2221 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     2281 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     2341 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     2401 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     2461 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     2521 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     2581 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     2641 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     2701 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     2761 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     2821 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     2881 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     2941 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     3001 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     3061 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     3121 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     3181 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     3241 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     3301 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     3361 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     3421 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     3481 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     3541 attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat ggagagtaaa
     3601 cggaataagc cagggaaggc gacaggtaaa ggtaaaccag ttggtgacaa gtggctggat
     3661 gatgcgggta aagactctgg tgcgccgatc ccagaccgta tcgcggacaa actgcgtgac
     3721 aaagaattca aaaacttcga cgacttccgt aagaaattct ggaaagaagt tgcgaaagac
     3781 ccggatctcg ctaaacagtt caaacgtgcg aaccgtcgta acatcaaaca gggtcgtgcg
     3841 ccattcgcgc cgaaaaaaga tcaggttggt ggtcgtcgta ccttcgaact gcaccacgac
     3901 aaaccgatct ctcaggacgg tggtgtttac gacatgaaca acatcagagt gaccacacct
     3961 aagcgacata ttgatattca tcggggtaag taaaaatgga actgaaacat agtattagtg
     4021 attataccga ggctgaattt ctggagtttg tgaagaaaat ctacaactct accgacgaag
     4081 agaaacagaa ggaactggtt accgaatttg aacgtctgac cgaacacccg tctggttctg
     4141 acctgatcta ctacccgcgt gacgaccgtg aagactctcc ggaaggtatc gttaaggaaa
     4201 tcaaggagtg gcgagctgct aacggtaagt caggatttaa acagggcctc gagcaccacc
     4261 accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     4321 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     4381 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
     4441 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4501 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4561 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4621 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4681 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4741 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4801 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4861 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4921 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4981 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5041 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5101 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5161 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5221 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5281 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5341 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5401 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5461 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5521 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5581 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5641 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5701 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5761 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5821 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5881 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5941 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6001 tgagcgtcag accccgtaga
//
