LOCUS       sequence-448051-        7514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpR and GvpF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  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 7514)
  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'-deltaRN
FEATURES             Location/Qualifiers
     source          1..7514
                     /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             2012..2029
                     /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(5772..5790)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5786..5833
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5900..5991
                     /gene="bla"
                     /label=AmpR promoter
     terminator      6856..6895
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      6926..7514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7415..7434
                     /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 ctttaggagc
     1441 ttgaaaacat gagtgaaaca aacgaaacag gtatttatat ttttagcgcc attcaaacgg
     1501 ataaagacga agaatttggc gccgtggaag tagaaggaac aaaagctgaa acatttttga
     1561 ttcgctacaa agacgcggct atggtagcag ctgaagtacc gatgaaaatt tatcatccta
     1621 atcgccaaaa tttattaatg catcaaaacg cagtagcagc gattatggac aagaacgata
     1681 cggttattcc aatcagcttt gggaatgtat tcaaatcaaa agaagacgta aaagttcttt
     1741 tggaaaacct ttatccgcag tttgaaaagc tgtttccagc gatcaaagga aaaattgaag
     1801 tcggtttaaa agtaattggg aaaaaagaat ggcttgagaa aaaagtaaac gaaaatcctg
     1861 aacttgagaa agtatcagca tccgtaaaag gaaaatcaga agcagccggt tattatgagc
     1921 gtattcaact tggaggaatg gctcaaaaga tgtttacttc cctgcaaaaa gaagtcaaga
     1981 cagatgtgtt ttctccgctt gaagaagcag cggaagcagc aaaagcaaat gagccaacgg
     2041 gcgaaacgat gcttttaaac gcgtctttct taattaaccg agaagatgaa gcgaagtttg
     2101 atgaaaaagt aaatgaagcg catgaaaact ggaaagacaa agccgatttt cattacagcg
     2161 gtccttggcc tgcttataat tttgtgaaca ttcgcctaaa agtagaagag aaataacgtg
     2221 cttcacaaat tagtaaccgc acccattaac cttgtagtga aaatcggcga aaaagtacag
     2281 gaagaagctg ataaacagct atatgacctt ccgacgattc agcaaaagct cattcagctt
     2341 caaatgatgt ttgagcttgg tgaaattcca gaagaagcgt ttcaagaaaa agaagatgaa
     2401 ttgttaatga ggtacgaaat tgcgaaacgc agagaaattg aacaatggga agagctaaca
     2461 caaaaaagaa atgaggaatc ctagatggga gaattactgt atttatacgg tttaattcca
     2521 acaaaagaag cagcagccat agagccgttt ccatcttata aggggtttga cggagaacat
     2581 tcactgtacc caattgcgtt tgatcaggtg acggctgtag tttctaagct ggatgctgac
     2641 acctattcag aaaaagtgat tcaagaaaaa atggagcagg atatgagctg gctgcaggaa
     2701 aaagcatttc atcatcacga aacggtagcc gctttgtacg aagaatttac gatcattcca
     2761 ttaaaatttt gcaccattta taaaggtgaa gaaagtctgc aggcagctat tgagattaac
     2821 aaagaaaaga tagagaattc actgacgctg cttcaaggaa atgaagagtg gaatgtgaaa
     2881 atttactgtg atgatacaga gcttaaaaaa ggaatcagcg aaacgaatga aagcgtgaaa
     2941 gcgaaaaaac aagaaattag tcacttatca ccaggaagac agttttttga aaagaaaaaa
     3001 atagatcagc tgattgaaaa agaattagag cttcacaaaa acaaagtgtg tgaagagata
     3061 catgacaagc taaaagaatt atcgctttat gactctgtta aaaagaattg gagcaaagac
     3121 gtaactggcg cagctgaaca gatggcgtgg aacagcgtgt ttcttctccc gtctctgcag
     3181 attactaagt tcgtaaacga aatagaagag cttcagcaaa ggcttgaaaa taaaggctgg
     3241 aagtttgaag tgacgggacc atggccgccc tatcatttct cgagctttgc gtaaagtgag
     3301 gaattaacat tatgtctctt aaacaatcca tggagaataa agatattgct cttattgata
     3361 ttttagatgt cattttagat aaaggagtcg ccattaaagg agacttaatc atttccatag
     3421 ctggcgtcga tttagtgtat ttggatttgc gggtgcttat ttcttcggtt gaaacgcttg
     3481 tgcaagcaaa agaaggaaat cacaaaccaa tcacttctga acaatttgat aaacaaaagg
     3541 aggaattaat ggatgcaacc ggtcagccaa gcaaatggac gaatccactt ggatcctgat
     3601 caagctgaac aaggcttagc gcagcttgtg atgacagtta ttgagctatt gaggcaaatc
     3661 gttgaacgtc atgccatgag gcgggtggag ggtggaacgt tgacggacga acaaattgaa
     3721 aacttaggaa ttgcactaat gaacttagaa gaaaaaatgg acgagttgaa agaggtgttc
     3781 ggtctggatg cagaagattt aaatattgat cttggaccgc taggcagcct gctttaagcg
     3841 gtcagtagga ggaacagtat ggcagtcgaa cataatatgc agtcaagtac gattgtagat
     3901 gtgctcgaaa agattttgga taaaggagtc gttatagcgg gggacatcac cgtaggaatt
     3961 gcagatgtcg agctattaac gataaagatc cgcttgattg tggcttcggt tgataaggca
     4021 aaagaaatcg gcatggactg gtgggaaaat gatccgtatc tcagttcaaa aggagccaat
     4081 aacaaagcgc tcgaagaaga aaataaaatg ctgcatgagc ggttaaaaac gcttgaagaa
     4141 aaaatagaaa cgaaacgtta aaaactgtac gctacttaaa aaatggaggg atttacaatg
     4201 gcaactgaaa caaaattaga taacacacag gcagaaaaca aggaaaataa aaatgcggaa
     4261 aacggttcaa aagaaaagaa cggttcaaaa gcaagcaaaa caacaagcag cgggccaatc
     4321 aaacgagcgg tagcaggagg catcatcggt gcaacgattg gatatgtatc gactcctgaa
     4381 aatcgaaaaa gtctccttga ccgcattgat acagacgaat taaaaagcaa agcatctgat
     4441 ttaggaacaa aggtaaaaga aaaatcaaaa agcagcgtgg ccagcctgaa aacatctgcg
     4501 ggaagcttgt ttaaaaaaga taaagataaa tcaaaagatg atgaagaaaa cgtaaattct
     4561 tctagtagcg aaacagaaga cgataacgtt caagagtacg acgagttaaa agaagaaaat
     4621 caaactcttc aagatcgctt atcacagctt gaagaaaaaa tgaacatgct tgttgagctt
     4681 agcctcaata aaaatcaaga cgaagaagcg gaagatacag attccgacga agaagagaac
     4741 gatgagaacg atgaaaacga tgaaaacgag caggacgatg aaaacgaaga agaaacatct
     4801 aagccacgta aaaaggataa aaaagaagct gaggaagaag aaagtgaaag tgacgaagac
     4861 agcgaggaag aagaggaaga ttctcgctca aacaaaaaaa ataaaaaagt aaaaacagaa
     4921 gaagaagacg aagatgaaag cgaagaagaa aaaaaggaag cgaaaccaaa aaagtcaaca
     4981 gctaaaaaat caaaaaatac aaaagcaaag aaaaacacgg acgaagaaga tgatgaagca
     5041 acatctcttt ctagtgaaga cgatacaaca gcctaagacg taaaggagga aagaaagaca
     5101 tgagtacagg cccttctttt tcaactaaag acaatacgct tgaatacttt gtgaaagctt
     5161 ctaataaaca cggcttttca cttgatattt cattaaatgt aaacggcgct gtgatttccg
     5221 gtaccatgat ttcagcaaaa gaatactttg attacttaag cgaaacgttt gaagaaggca
     5281 gtgaagtggc tcaggcgcta agcgaacaat tctctttagc aagcgaagcg agcgaatcaa
     5341 acggagaagc agaagcccat tttattcatt tgaaaaatac aaagatttac tgtggagaca
     5401 gtaaatctac tccttctaaa ggcaaaatct tttggagagg gaaaatagca gaagtagacg
     5461 ggtttttctt aggaaagatt tctgatgcaa aatcaacgag taaaaagagt tcataaaaaa
     5521 cggcggggtg attgccccgc cgttttttag tgatgtgatg agatgtgcag cttctttttt
     5581 cgatacatga gcggagtcat cttcatttct tttcgaaact gattaataaa atagcttgta
     5641 ctattaaacc ctacttgata ggcgacttcc gtcacatttg cttctgcttg ctgcagtccg
     5701 gattcgcgaa cgcgtctcga gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     5761 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     5821 gaggggtttt ttgctgaaag gaggaactat atccgactag tagcccgcct aatgagcggg
     5881 ctttttttta attcccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6061 cacccagaaa cgctcgtgaa agtaaaagac gcagaggacc aattgggggc acgagtggga
     6121 tacatagaac tggacttgaa tagcggtaaa atccttgaga gttttcgccc tgaagagcgt
     6181 tttccaatga tgagcacttt caaagttctg ctatgtggag cagtattatc ccgtgtagat
     6241 gcggggcaag agcaactcgg acgacgaata cactattcgc agaatgactt ggttgaatac
     6301 tccccagtga cagaaaagca ccttacggac ggaatgacgg taagagaatt atgtagtgcc
     6361 gccataacga tgagtgataa cactgcggcg aacttacttc tgacaaccat cggtggaccg
     6421 aaggaattaa ccgctttttt gcacaatatg ggagaccatg taactcgcct tgaccgttgg
     6481 gaaccagaac tgaatgaagc cataccaaac gacgagcgag acaccacaat gcctgcggca
     6541 atggcaacaa cattacgcaa actattaact ggcgaactac ttactctggc ttcacggcaa
     6601 caattaatag actggcttga agcggataaa gttgcaggac cactactgcg ttcggcactt
     6661 cctgctggct ggtttattgc tgataaatct ggggcaggag agcgtggttc acggggtatc
     6721 attgccgcac ttggaccaga tggtaagcct tcccgtatcg tagttatcta cacgacgggt
     6781 agtcaggcaa ctatggacga acgaaataga cagattgctg aaataggggc ttcactgatt
     6841 aagcattggt aaaccgatac aattaaaggc tccttttgga gccttttttt ttggacggac
     6901 cggtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6961 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7021 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     7081 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7141 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7201 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7261 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7321 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7381 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7441 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7501 ggagcctatg gaaa
//
