LOCUS       sequence-446041-        8313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpR and GvpK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8313)
  AUTHORS   Iburg M, Anderson AP, Wong VT, Anton ED, He A, Lu GJ
  TITLE     Elucidating the assembly of gas vesicles by systematic 
            protein-protein interaction analysis.
  JOURNAL   EMBO J. 2024 Oct;43(19):4156-4172. doi: 10.1038/s44318-024-00178-2. 
            Epub 2024 Sep 3.
  PUBMED    39227754
REFERENCE   2  (bases 1 to 8313)
  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"; doi: 
            "10.1038/s44318-024-00178-2"; journalName: "EMBO J"; date: 
            "2024-10"; volume: "43"; issue: "19"; pages: "4156-4172"
COMMENT     Sequence Label: pST39-Mega'-deltaRK
FEATURES             Location/Qualifiers
     source          1..8313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    186..325
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(251..270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     411..433
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(427..618)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     855..874
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        855..873
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2996..3013
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="GSSKSK"
     primer_bind     complement(6571..6589)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6585..6632
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6699..6790
                     /gene="bla"
                     /label=AmpR promoter
     terminator      7655..7694
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      7725..8313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8214..8233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
      241 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      301 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      361 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      421 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
      481 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
      541 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
      601 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tggggtaatg
      661 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
      721 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
      781 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttgg caccatgcat
      841 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaga
      901 tagcttagcg atgtgaaaga agctggagaa aactttcaag aaaaagtttc ttcttcagac
      961 gataaagaga agtcaaaaaa taagcggaaa ataaaaggcg taaatcagat taaaaaagca
     1021 acggatatta aaagctctac taagattaaa agttctaatg atattaaatc atctacagat
     1081 ttaaaaacaa tgggatcata agcgaaagga gaaatgagat atgtctattc aaaaaagtac
     1141 taatagttca agtttagcag aagtcattga ccgtatttta gataaaggaa ttgttattga
     1201 tgcttttgca agagtttctg ttgtaggaat tgaaatttta acgattgaag cgcgagtggt
     1261 tattgccagt gttgatacat ggttacgcta tgcagaagca gtagggcttc ttcgtgacga
     1321 cgtagaagaa aacggtcttc ctgaacgttc aaattcaagt gaagggcagc cgcgttttag
     1381 tatttaatat gaagtggtat ggaaccttcc ttttttgagg gtggttccag cttttagatc
     1441 taactattgg aggctactaa aaatggaaaa gatggcagga ggaacgtaaa aatgaccgtc
     1501 ttaacagaca aaaggaaaaa aggcagtgga gcttttatac aagatgacga gacaaaagag
     1561 gttctttcaa gagcgctgag ctatttaaaa tccggctatt ccattcattt tacaggtcct
     1621 gccggcggag gcaaaacctc tttagcgcga gcgcttgcta aaaagagaaa gcgtcctgta
     1681 atgctgatgc acgggaatca cgagctcaac aacaaagatt taattggcga ttttacggga
     1741 tacacgagca aaaaagtaat cgaccagtac gttcgttctg tctataaaaa agatgaacag
     1801 gtgagtgaaa actggcagga tggccgattg cttgaagctg taaaaaatgg ctatacgctg
     1861 atttacgacg aatttactcg ttctaagcct gcgacgaata atatctttct atcgatatta
     1921 gaagaaggcg tgctgccgct gtatggagta aaaatgaccg atccttttgt gcgcgtgcat
     1981 cccgatttcc gcgtcatctt cacaagcaat ccagctgagt atgccggcgt atatgatacg
     2041 caagatgcgc ttctcgacag gttaattacc atgtttattg attataaaga catcgacaga
     2101 gagacagcga ttttaacgga gaaaacggac gtagaagaag atgaagcgcg cacaattgta
     2161 acgctcgtag caaacgtgcg aaaccgctct ggagacgaaa acagcagcgg acttagcctg
     2221 cgggcttcgc ttatgatcgc tacccttgcc acgcagcaag acattcctat cgatggaagt
     2281 gacgaagatt ttcaaacgtt atgtatcgat attttgcatc atccgcttac caaatgtttg
     2341 gatgaagaaa atgcaaaaag caaagccgaa aaaatcattt tagaagaatg taagaatata
     2401 gacactgaag aaaagtaaag gagcttgaaa acatgagtga aacaaacgaa acaggtattt
     2461 atatttttag cgccattcaa acggataaag acgaagaatt tggcgccgtg gaagtagaag
     2521 gaacaaaagc tgaaacattt ttgattcgct acaaagacgc ggctatggta gcagctgaag
     2581 taccgatgaa aatttatcat cctaatcgcc aaaatttatt aatgcatcaa aacgcagtag
     2641 cagcgattat ggacaagaac gatacggtta ttccaatcag ctttgggaat gtattcaaat
     2701 caaaagaaga cgtaaaagtt cttttggaaa acctttatcc gcagtttgaa aagctgtttc
     2761 cagcgatcaa aggaaaaatt gaagtcggtt taaaagtaat tgggaaaaaa gaatggcttg
     2821 agaaaaaagt aaacgaaaat cctgaacttg agaaagtatc agcatccgta aaaggaaaat
     2881 cagaagcagc cggttattat gagcgtattc aacttggagg aatggctcaa aagatgttta
     2941 cttccctgca aaaagaagtc aagacagatg tgttttctcc gcttgaagaa gcagcggaag
     3001 cagcaaaagc aaatgagcca acgggcgaaa cgatgctttt aaacgcgtct ttcttaatta
     3061 accgagaaga tgaagcgaag tttgatgaaa aagtaaatga agcgcatgaa aactggaaag
     3121 acaaagccga ttttcattac agcggtcctt ggcctgctta taattttgtg aacattcgcc
     3181 taaaagtaga agagaaataa cgtgcttcac aaattagtaa ccgcacccat taaccttgta
     3241 gtgaaaatcg gcgaaaaagt acaggaagaa gctgataaac agctatatga ccttccgacg
     3301 attcagcaaa agctcattca gcttcaaatg atgtttgagc ttggtgaaat tccagaagaa
     3361 gcgtttcaag aaaaagaaga tgaattgtta atgaggtacg aaattgcgaa acgcagagaa
     3421 attgaacaat gggaagagct aacacaaaaa agaaatgagg aatcctagat gggagaatta
     3481 ctgtatttat acggtttaat tccaacaaaa gaagcagcag ccatagagcc gtttccatct
     3541 tataaggggt ttgacggaga acattcactg tacccaattg cgtttgatca ggtgacggct
     3601 gtagtttcta agctggatgc tgacacctat tcagaaaaag tgattcaaga aaaaatggag
     3661 caggatatga gctggctgca ggaaaaagca tttcatcatc acgaaacggt agccgctttg
     3721 tacgaagaat ttacgatcat tccattaaaa ttttgcacca tttataaagg tgaagaaagt
     3781 ctgcaggcag ctattgagat taacaaagaa aagatagaga attcactgac gctgcttcaa
     3841 ggaaatgaag agtggaatgt gaaaatttac tgtgatgata cagagcttaa aaaaggaatc
     3901 agcgaaacga atgaaagcgt gaaagcgaaa aaacaagaaa ttagtcactt atcaccagga
     3961 agacagtttt ttgaaaagaa aaaaatagat cagctgattg aaaaagaatt agagcttcac
     4021 aaaaacaaag tgtgtgaaga gatacatgac aagctaaaag aattatcgct ttatgactct
     4081 gttaaaaaga attggagcaa agacgtaact ggcgcagctg aacagatggc gtggaacagc
     4141 gtgtttcttc tcccgtctct gcagattact aagttcgtaa acgaaataga agagcttcag
     4201 caaaggcttg aaaataaagg ctggaagttt gaagtgacgg gaccatggcc gccctatcat
     4261 ttctcgagct ttgcgtaaag tgaggaatta acattatgtc tcttaaacaa tccatggaga
     4321 ataaagatat tgctcttatt gatattttag atgtcatttt agataaagga gtcgccatta
     4381 aaggagactt aatcatttcc atagctggcg tcgatttagt gtatttggat ttgcgggtgc
     4441 ttatttcttc ggttgaaacg cttgtgcaag caaaagaagg aaatcacaaa ccaatcactt
     4501 ctgaacaatt tgataaacaa aaagaagagc tgatggacgc tacgggccag cctagtaagt
     4561 ggacgaaccc tctaggttcg tgacccttgc gagcagtcgt atgctttctc gaattccgag
     4621 cggttaagcg tgacaggcgg tcagtaggag gaacagtatg gcagtcgaac ataatatgca
     4681 gtcaagtacg attgtagatg tgctcgaaaa gattttggat aaaggagtcg ttatagcggg
     4741 ggacatcacc gtaggaattg cagatgtcga gctattaacg ataaagatcc gcttgattgt
     4801 ggcttcggtt gataaggcaa aagaaatcgg catggactgg tgggaaaatg atccgtatct
     4861 cagttcaaaa ggagccaata acaaagcgct cgaagaagaa aataaaatgc tgcatgagcg
     4921 gttaaaaacg cttgaagaaa aaatagaaac gaaacgttaa aaactgtacg ctacttaaaa
     4981 aatggaggga tttacaatgg caactgaaac aaaattagat aacacacagg cagaaaacaa
     5041 ggaaaataaa aatgcggaaa acggttcaaa agaaaagaac ggttcaaaag caagcaaaac
     5101 aacaagcagc gggccaatca aacgagcggt agcaggaggc atcatcggtg caacgattgg
     5161 atatgtatcg actcctgaaa atcgaaaaag tctccttgac cgcattgata cagacgaatt
     5221 aaaaagcaaa gcatctgatt taggaacaaa ggtaaaagaa aaatcaaaaa gcagcgtggc
     5281 cagcctgaaa acatctgcgg gaagcttgtt taaaaaagat aaagataaat caaaagatga
     5341 tgaagaaaac gtaaattctt ctagtagcga aacagaagac gataacgttc aagagtacga
     5401 cgagttaaaa gaagaaaatc aaactcttca agatcgctta tcacagcttg aagaaaaaat
     5461 gaacatgctt gttgagctta gcctcaataa aaatcaagac gaagaagcgg aagatacaga
     5521 ttccgacgaa gaagagaacg atgagaacga tgaaaacgat gaaaacgagc aggacgatga
     5581 aaacgaagaa gaaacatcta agccacgtaa aaaggataaa aaagaagctg aggaagaaga
     5641 aagtgaaagt gacgaagaca gcgaggaaga agaggaagat tctcgctcaa acaaaaaaaa
     5701 taaaaaagta aaaacagaag aagaagacga agatgaaagc gaagaagaaa aaaaggaagc
     5761 gaaaccaaaa aagtcaacag ctaaaaaatc aaaaaataca aaagcaaaga aaaacacgga
     5821 cgaagaagat gatgaagcaa catctctttc tagtgaagac gatacaacag cctaagacgt
     5881 aaaggaggaa agaaagacat gagtacaggc ccttcttttt caactaaaga caatacgctt
     5941 gaatactttg tgaaagcttc taataaacac ggcttttcac ttgatatttc attaaatgta
     6001 aacggcgctg tgatttccgg taccatgatt tcagcaaaag aatactttga ttacttaagc
     6061 gaaacgtttg aagaaggcag tgaagtggct caggcgctaa gcgaacaatt ctctttagca
     6121 agcgaagcga gcgaatcaaa cggagaagca gaagcccatt ttattcattt gaaaaataca
     6181 aagatttact gtggagacag taaatctact ccttctaaag gcaaaatctt ttggagaggg
     6241 aaaatagcag aagtagacgg gtttttctta ggaaagattt ctgatgcaaa atcaacgagt
     6301 aaaaagagtt cataaaaaac ggcggggtga ttgccccgcc gttttttagt gatgtgatga
     6361 gatgtgcagc ttcttttttc gatacatgag cggagtcatc ttcatttctt ttcgaaactg
     6421 attaataaaa tagcttgtac tattaaaccc tacttgatag gcgacttccg tcacatttgc
     6481 ttctgcttgc tgcagtccgg attcgcgaac gcgtctcgag agatccggct gctaacaaag
     6541 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     6601 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccgactagt
     6661 agcccgccta atgagcgggc ttttttttaa ttcccctatt tgtttatttt tctaaataca
     6721 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6781 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6841 ttgccttcct gtttttgctc acccagaaac gctcgtgaaa gtaaaagacg cagaggacca
     6901 attgggggca cgagtgggat acatagaact ggacttgaat agcggtaaaa tccttgagag
     6961 ttttcgccct gaagagcgtt ttccaatgat gagcactttc aaagttctgc tatgtggagc
     7021 agtattatcc cgtgtagatg cggggcaaga gcaactcgga cgacgaatac actattcgca
     7081 gaatgacttg gttgaatact ccccagtgac agaaaagcac cttacggacg gaatgacggt
     7141 aagagaatta tgtagtgccg ccataacgat gagtgataac actgcggcga acttacttct
     7201 gacaaccatc ggtggaccga aggaattaac cgcttttttg cacaatatgg gagaccatgt
     7261 aactcgcctt gaccgttggg aaccagaact gaatgaagcc ataccaaacg acgagcgaga
     7321 caccacaatg cctgcggcaa tggcaacaac attacgcaaa ctattaactg gcgaactact
     7381 tactctggct tcacggcaac aattaataga ctggcttgaa gcggataaag ttgcaggacc
     7441 actactgcgt tcggcacttc ctgctggctg gtttattgct gataaatctg gggcaggaga
     7501 gcgtggttca cggggtatca ttgccgcact tggaccagat ggtaagcctt cccgtatcgt
     7561 agttatctac acgacgggta gtcaggcaac tatggacgaa cgaaatagac agattgctga
     7621 aataggggct tcactgatta agcattggta aaccgataca attaaaggct ccttttggag
     7681 cctttttttt tggacggacc ggtagaaaag atcaaaggat cttcttgaga tccttttttt
     7741 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7801 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7861 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7921 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7981 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8041 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8101 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8161 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8221 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8281 tgatgctcgt caggggggcg gagcctatgg aaa
//
