LOCUS       sequence-445987-        8236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpR and GvpS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8236)
  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 8236)
  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'-deltaRS
FEATURES             Location/Qualifiers
     source          1..8236
                     /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             2955..2972
                     /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(6494..6512)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6508..6555
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6622..6713
                     /gene="bla"
                     /label=AmpR promoter
     terminator      7578..7617
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      7648..8236
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8137..8156
                     /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 ctttcaggag
     1441 gaacgtaaaa atgaccgtct taacagacaa aaggaaaaaa ggcagtggag cttttataca
     1501 agatgacgag acaaaagagg ttctttcaag agcgctgagc tatttaaaat ccggctattc
     1561 cattcatttt acaggtcctg ccggcggagg caaaacctct ttagcgcgag cgcttgctaa
     1621 aaagagaaag cgtcctgtaa tgctgatgca cgggaatcac gagctcaaca acaaagattt
     1681 aattggcgat tttacgggat acacgagcaa aaaagtaatc gaccagtacg ttcgttctgt
     1741 ctataaaaaa gatgaacagg tgagtgaaaa ctggcaggat ggccgattgc ttgaagctgt
     1801 aaaaaatggc tatacgctga tttacgacga atttactcgt tctaagcctg cgacgaataa
     1861 tatctttcta tcgatattag aagaaggcgt gctgccgctg tatggagtaa aaatgaccga
     1921 tccttttgtg cgcgtgcatc ccgatttccg cgtcatcttc acaagcaatc cagctgagta
     1981 tgccggcgta tatgatacgc aagatgcgct tctcgacagg ttaattacca tgtttattga
     2041 ttataaagac atcgacagag agacagcgat tttaacggag aaaacggacg tagaagaaga
     2101 tgaagcgcgc acaattgtaa cgctcgtagc aaacgtgcga aaccgctctg gagacgaaaa
     2161 cagcagcgga cttagcctgc gggcttcgct tatgatcgct acccttgcca cgcagcaaga
     2221 cattcctatc gatggaagtg acgaagattt tcaaacgtta tgtatcgata ttttgcatca
     2281 tccgcttacc aaatgtttgg atgaagaaaa tgcaaaaagc aaagccgaaa aaatcatttt
     2341 agaagaatgt aagaatatag acactgaaga aaagtaaagg agcttgaaaa catgagtgaa
     2401 acaaacgaaa caggtattta tatttttagc gccattcaaa cggataaaga cgaagaattt
     2461 ggcgccgtgg aagtagaagg aacaaaagct gaaacatttt tgattcgcta caaagacgcg
     2521 gctatggtag cagctgaagt accgatgaaa atttatcatc ctaatcgcca aaatttatta
     2581 atgcatcaaa acgcagtagc agcgattatg gacaagaacg atacggttat tccaatcagc
     2641 tttgggaatg tattcaaatc aaaagaagac gtaaaagttc ttttggaaaa cctttatccg
     2701 cagtttgaaa agctgtttcc agcgatcaaa ggaaaaattg aagtcggttt aaaagtaatt
     2761 gggaaaaaag aatggcttga gaaaaaagta aacgaaaatc ctgaacttga gaaagtatca
     2821 gcatccgtaa aaggaaaatc agaagcagcc ggttattatg agcgtattca acttggagga
     2881 atggctcaaa agatgtttac ttccctgcaa aaagaagtca agacagatgt gttttctccg
     2941 cttgaagaag cagcggaagc agcaaaagca aatgagccaa cgggcgaaac gatgctttta
     3001 aacgcgtctt tcttaattaa ccgagaagat gaagcgaagt ttgatgaaaa agtaaatgaa
     3061 gcgcatgaaa actggaaaga caaagccgat tttcattaca gcggtccttg gcctgcttat
     3121 aattttgtga acattcgcct aaaagtagaa gagaaataac gtgcttcaca aattagtaac
     3181 cgcacccatt aaccttgtag tgaaaatcgg cgaaaaagta caggaagaag ctgataaaca
     3241 gctatatgac cttccgacga ttcagcaaaa gctcattcag cttcaaatga tgtttgagct
     3301 tggtgaaatt ccagaagaag cgtttcaaga aaaagaagat gaattgttaa tgaggtacga
     3361 aattgcgaaa cgcagagaaa ttgaacaatg ggaagagcta acacaaaaaa gaaatgagga
     3421 atcctagatg ggagaattac tgtatttata cggtttaatt ccaacaaaag aagcagcagc
     3481 catagagccg tttccatctt ataaggggtt tgacggagaa cattcactgt acccaattgc
     3541 gtttgatcag gtgacggctg tagtttctaa gctggatgct gacacctatt cagaaaaagt
     3601 gattcaagaa aaaatggagc aggatatgag ctggctgcag gaaaaagcat ttcatcatca
     3661 cgaaacggta gccgctttgt acgaagaatt tacgatcatt ccattaaaat tttgcaccat
     3721 ttataaaggt gaagaaagtc tgcaggcagc tattgagatt aacaaagaaa agatagagaa
     3781 ttcactgacg ctgcttcaag gaaatgaaga gtggaatgtg aaaatttact gtgatgatac
     3841 agagcttaaa aaaggaatca gcgaaacgaa tgaaagcgtg aaagcgaaaa aacaagaaat
     3901 tagtcactta tcaccaggaa gacagttttt tgaaaagaaa aaaatagatc agctgattga
     3961 aaaagaatta gagcttcaca aaaacaaagt gtgtgaagag atacatgaca agctaaaaga
     4021 attatcgctt tatgactctg ttaaaaagaa ttggagcaaa gacgtaactg gcgcagctga
     4081 acagatggcg tggaacagcg tgtttcttct cccgtctctg cagattacta agttcgtaaa
     4141 cgaaatagaa gagcttcagc aaaggcttga aaataaaggc tggaagtttg aagtgacggg
     4201 accatggccg ccctatcatt tctcgagctt tccgtaaaag cggttaagcg tgacagtcca
     4261 ggaggaatta atggatgcaa ccggtcagcc aagcaaatgg acgaatccac ttggatcctg
     4321 atcaagctga acaaggctta gcgcagcttg tgatgacagt tattgagcta ttgaggcaaa
     4381 tcgttgaacg tcatgccatg aggcgggtgg agggtggaac gttgacggac gaacaaattg
     4441 aaaacttagg aattgcacta atgaacttag aagaaaaaat ggacgagttg aaagaggtgt
     4501 tcggtctgga tgcagaagat ttaaatattg atcttggacc gctaggcagc ctgctttaag
     4561 cggtcagtag gaggaacagt atggcagtcg aacataatat gcagtcaagt acgattgtag
     4621 atgtgctcga aaagattttg gataaaggag tcgttatagc gggggacatc accgtaggaa
     4681 ttgcagatgt cgagctatta acgataaaga tccgcttgat tgtggcttcg gttgataagg
     4741 caaaagaaat cggcatggac tggtgggaaa atgatccgta tctcagttca aaaggagcca
     4801 ataacaaagc gctcgaagaa gaaaataaaa tgctgcatga gcggttaaaa acgcttgaag
     4861 aaaaaataga aacgaaacgt taaaaactgt acgctactta aaaaatggag ggatttacaa
     4921 tggcaactga aacaaaatta gataacacac aggcagaaaa caaggaaaat aaaaatgcgg
     4981 aaaacggttc aaaagaaaag aacggttcaa aagcaagcaa aacaacaagc agcgggccaa
     5041 tcaaacgagc ggtagcagga ggcatcatcg gtgcaacgat tggatatgta tcgactcctg
     5101 aaaatcgaaa aagtctcctt gaccgcattg atacagacga attaaaaagc aaagcatctg
     5161 atttaggaac aaaggtaaaa gaaaaatcaa aaagcagcgt ggccagcctg aaaacatctg
     5221 cgggaagctt gtttaaaaaa gataaagata aatcaaaaga tgatgaagaa aacgtaaatt
     5281 cttctagtag cgaaacagaa gacgataacg ttcaagagta cgacgagtta aaagaagaaa
     5341 atcaaactct tcaagatcgc ttatcacagc ttgaagaaaa aatgaacatg cttgttgagc
     5401 ttagcctcaa taaaaatcaa gacgaagaag cggaagatac agattccgac gaagaagaga
     5461 acgatgagaa cgatgaaaac gatgaaaacg agcaggacga tgaaaacgaa gaagaaacat
     5521 ctaagccacg taaaaaggat aaaaaagaag ctgaggaaga agaaagtgaa agtgacgaag
     5581 acagcgagga agaagaggaa gattctcgct caaacaaaaa aaataaaaaa gtaaaaacag
     5641 aagaagaaga cgaagatgaa agcgaagaag aaaaaaagga agcgaaacca aaaaagtcaa
     5701 cagctaaaaa atcaaaaaat acaaaagcaa agaaaaacac ggacgaagaa gatgatgaag
     5761 caacatctct ttctagtgaa gacgatacaa cagcctaaga cgtaaaggag gaaagaaaga
     5821 catgagtaca ggcccttctt tttcaactaa agacaatacg cttgaatact ttgtgaaagc
     5881 ttctaataaa cacggctttt cacttgatat ttcattaaat gtaaacggcg ctgtgatttc
     5941 cggtaccatg atttcagcaa aagaatactt tgattactta agcgaaacgt ttgaagaagg
     6001 cagtgaagtg gctcaggcgc taagcgaaca attctcttta gcaagcgaag cgagcgaatc
     6061 aaacggagaa gcagaagccc attttattca tttgaaaaat acaaagattt actgtggaga
     6121 cagtaaatct actccttcta aaggcaaaat cttttggaga gggaaaatag cagaagtaga
     6181 cgggtttttc ttaggaaaga tttctgatgc aaaatcaacg agtaaaaaga gttcataaaa
     6241 aacggcgggg tgattgcccc gccgtttttt agtgatgtga tgagatgtgc agcttctttt
     6301 ttcgatacat gagcggagtc atcttcattt cttttcgaaa ctgattaata aaatagcttg
     6361 tactattaaa ccctacttga taggcgactt ccgtcacatt tgcttctgct tgctgcagtc
     6421 cggattcgcg aacgcgtctc gagagatccg gctgctaaca aagcccgaaa ggaagctgag
     6481 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     6541 ttgaggggtt ttttgctgaa aggaggaact atatccgact agtagcccgc ctaatgagcg
     6601 ggcttttttt taattcccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6781 ctcacccaga aacgctcgtg aaagtaaaag acgcagagga ccaattgggg gcacgagtgg
     6841 gatacataga actggacttg aatagcggta aaatccttga gagttttcgc cctgaagagc
     6901 gttttccaat gatgagcact ttcaaagttc tgctatgtgg agcagtatta tcccgtgtag
     6961 atgcggggca agagcaactc ggacgacgaa tacactattc gcagaatgac ttggttgaat
     7021 actccccagt gacagaaaag caccttacgg acggaatgac ggtaagagaa ttatgtagtg
     7081 ccgccataac gatgagtgat aacactgcgg cgaacttact tctgacaacc atcggtggac
     7141 cgaaggaatt aaccgctttt ttgcacaata tgggagacca tgtaactcgc cttgaccgtt
     7201 gggaaccaga actgaatgaa gccataccaa acgacgagcg agacaccaca atgcctgcgg
     7261 caatggcaac aacattacgc aaactattaa ctggcgaact acttactctg gcttcacggc
     7321 aacaattaat agactggctt gaagcggata aagttgcagg accactactg cgttcggcac
     7381 ttcctgctgg ctggtttatt gctgataaat ctggggcagg agagcgtggt tcacggggta
     7441 tcattgccgc acttggacca gatggtaagc cttcccgtat cgtagttatc tacacgacgg
     7501 gtagtcaggc aactatggac gaacgaaata gacagattgc tgaaataggg gcttcactga
     7561 ttaagcattg gtaaaccgat acaattaaag gctccttttg gagccttttt ttttggacgg
     7621 accggtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7681 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7741 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     7801 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7861 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7921 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7981 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8041 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     8101 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     8161 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8221 gcggagccta tggaaa
//
