LOCUS       sequence-204607-        6023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for expression of computationally designed colicin 
            endonuclease and immunity pair, des7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6023)
  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 6023)
  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-colEdes7/Imdes7
FEATURES             Location/Qualifiers
     source          1..6023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      21..609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     510..529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     763..780
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    795..934
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(860..879)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1020..1042
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1036..1227)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3079..3098
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3118..3195)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3401..3420
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3504..3523
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3504..3522
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3523..3547
                     /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             3562..3584
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4259..4276
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4329..4347)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4343..4390
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4427..4882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4514..4533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4724..4745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4908..5012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5013..5081,5082..5873)
                     /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(5231..5250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
       61 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
      121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
      181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
      241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
      301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
      361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
      421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
      481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
      541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
      601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
      721 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      781 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
      841 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
      901 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
      961 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     1021 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     1081 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     1141 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     1201 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     1261 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     1321 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     1381 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     1441 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     1501 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     1561 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     1621 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     1681 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     1741 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     1801 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     1861 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     1921 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     1981 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     2041 gcccgctttc cagtcgggaa actgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2101 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     2161 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     2221 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     2281 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     2341 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2401 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2461 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2521 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2581 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2641 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2701 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2761 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2821 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2881 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     2941 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     3001 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     3061 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     3121 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     3181 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     3241 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     3301 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     3361 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     3421 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     3481 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     3541 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atggagagta
     3601 aacggaataa gccagggaag gcgacaggta aaggtaaacc agttggtgac aagtggctgg
     3661 acgacgcggg taaagactct ggtgcgccga tcccggatcg tatcgcggac aagctccgtg
     3721 acaaagaatt caaaaacttc gacgacttcc gcaagaaatt ctgggaagaa gtttctaaag
     3781 acccggacct ggcgaaacag ttcaaacgtt ctaaccgtaa acgtatccag cagggttacg
     3841 cgccgttcgc gccacagaaa gaccaggttg gtggtcgtac caccttcgaa ctgcaccacg
     3901 acaaaccgat ctctcaagac ggtggtgttt acgatatgaa caacatcaga gtgaccacac
     3961 ctaagcgaca tattgatatt catcggggta agtaaaaatg gaactgaaac atagtattag
     4021 tgattatacc gaggctgaat ttctggagtt tgtgaagaaa atcgaagacg cgaactcttc
     4081 tgaagacgaa cagcagaaac tggttgaaga attcatccgt ctgaccgaac acccgtctgg
     4141 ttctgacctg atctactatc cgcgtgacga ccgtgaagac tctccggaag gtatcgttaa
     4201 agaaatcaaa gaatggcgag ctgctaacgg taagtcagga tttaaacagg gcctcgagca
     4261 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     4321 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     4381 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     4441 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4501 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4561 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4621 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4681 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4741 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4801 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4861 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4921 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4981 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5041 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5101 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5161 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5221 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5281 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5341 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5401 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5461 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5521 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     5581 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5641 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5701 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5761 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5821 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5881 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5941 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6001 tccactgagc gtcagacccc gta
//
