LOCUS       sequence-204353-        6032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for expression of computationally designed colicin 
            endonuclease and immunity pair, des12.
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-colEdes12/Imdes12
FEATURES             Location/Qualifiers
     source          1..6032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      24..612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     513..532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     766..783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    798..937
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(863..882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1023..1045
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1039..1230)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3082..3101
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3121..3198)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3404..3423
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3507..3526
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3507..3525
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3526..3550
                     /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             3565..3587
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4271..4288
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4341..4359)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4355..4402
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4439..4894
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4526..4545)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4736..4757
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4920..5024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5025..5093,5094..5885)
                     /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(5243..5262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
       61 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
      181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      661 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      721 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      781 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      841 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
      901 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
      961 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     1021 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     1081 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     1141 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     1201 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     1261 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     1321 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     1381 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     1441 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     1501 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     1561 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     1621 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     1681 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     1741 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     1801 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     1861 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     1921 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     1981 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     2041 actgcccgct ttccagtcgg gaaactgtcg tgccagctgc attaatgaat cggccaacgc
     2101 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     2161 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     2221 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     2281 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     2341 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     2401 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     2461 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     2521 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     2581 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     2641 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     2701 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     2761 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     2821 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     2881 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     2941 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     3001 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     3061 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     3121 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     3181 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     3241 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     3301 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     3361 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     3421 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     3481 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     3541 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatggagt
     3601 ctaaacgtaa caaaccgggt aaagccactg gcaaaggtaa acctgtcggc gacaaatggc
     3661 tggatgacgc cggtaaagac tctggtgcgc cgatcccaga ccgtatcgcg gacaaactgc
     3721 gtgacaaaga attcaaaaac ttcgacgact tcaagaaaaa gttttggaaa gaagtttcta
     3781 aagacccaga tctctctaag cagttcaaag gctctaaccg tgaaaacatc aagcagggta
     3841 ttgcgccgtt tgcgcgtcag aaagaccagg tcggtggtcg tcgtgttttc gaactgcacc
     3901 acgataaacc aatctctcag gacggtggtg tttacgacat gaacaacatc cgtgttacca
     3961 ccccgaaacg tcacatcgac atccaccggg gtaagtaaaa atggaactga aacactctat
     4021 ctctgactac accgaagcgg aatttctgga attcgttaag gaaatctgcc agaaaaactc
     4081 tgcgggtgaa ctggacgaat ctctgttcaa actggttacc gaatttgaac gtctgaccga
     4141 acacccggac ggttctgacc tgatctacta tccgcgtgac gatcgcgaag attctccgga
     4201 aggtatcgtt aaagaaatca aagagtggcg tgcggcgaac ggtaaatctg gtttcaaaca
     4261 aggcctcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     4321 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     4381 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     4441 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4501 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4561 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4621 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4681 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4741 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4801 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4861 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     4921 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4981 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5041 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5101 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5161 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5221 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5281 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5341 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5401 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5461 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5521 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     5581 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5641 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5701 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5761 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5821 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5881 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5941 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6001 gtgagttttc gttccactga gcgtcagacc cc
//
