LOCUS       sequence-445991-        8176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpU.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8176)
  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 8176)
  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'-deltaU
FEATURES             Location/Qualifiers
     source          1..8176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..121
                     /gene="bla"
                     /label=AmpR promoter
     terminator      986..1025
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1056..1644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1545..1564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1798..1815
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1830..1969
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1895..1914)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2055..2077
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2071..2262)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2499..2518
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2499..2517
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4906..4923
                     /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(8078..8096)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8092..8139
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 aatgagcggg ctttttttta attcccctat ttgtttattt ttctaaatac attcaaatat
       61 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      121 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      181 tgtttttgct cacccagaaa cgctcgtgaa agtaaaagac gcagaggacc aattgggggc
      241 acgagtggga tacatagaac tggacttgaa tagcggtaaa atccttgaga gttttcgccc
      301 tgaagagcgt tttccaatga tgagcacttt caaagttctg ctatgtggag cagtattatc
      361 ccgtgtagat gcggggcaag agcaactcgg acgacgaata cactattcgc agaatgactt
      421 ggttgaatac tccccagtga cagaaaagca ccttacggac ggaatgacgg taagagaatt
      481 atgtagtgcc gccataacga tgagtgataa cactgcggcg aacttacttc tgacaaccat
      541 cggtggaccg aaggaattaa ccgctttttt gcacaatatg ggagaccatg taactcgcct
      601 tgaccgttgg gaaccagaac tgaatgaagc cataccaaac gacgagcgag acaccacaat
      661 gcctgcggca atggcaacaa cattacgcaa actattaact ggcgaactac ttactctggc
      721 ttcacggcaa caattaatag actggcttga agcggataaa gttgcaggac cactactgcg
      781 ttcggcactt cctgctggct ggtttattgc tgataaatct ggggcaggag agcgtggttc
      841 acggggtatc attgccgcac ttggaccaga tggtaagcct tcccgtatcg tagttatcta
      901 cacgacgggt agtcaggcaa ctatggacga acgaaataga cagattgctg aaataggggc
      961 ttcactgatt aagcattggt aaaccgatac aattaaaggc tccttttgga gccttttttt
     1021 ttggacggac cggtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1081 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1141 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1201 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1261 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1321 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1381 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1441 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1501 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1561 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1621 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1681 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1741 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     1801 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     1861 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     1921 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     1981 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     2041 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     2101 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     2161 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     2221 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     2281 gggatttctg ttcatggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     2341 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     2401 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     2461 gatgtaggtg ttggcaccat gcatcgatcc cgcgaaatta atacgactca ctatagggag
     2521 accacaacgg tttccctcta gagatagctt agcgatgtga aagaagctgg agaaaacttt
     2581 caagaaaaag tttcttcttc agacgataaa gagaagtcaa aaaataagcg gaaaataaaa
     2641 ggcgtaaatc agattaaaaa agcaacggat attaaaagct ctactaagat taaaagttct
     2701 aatgatatta aatcatctac agatttaaaa acaatgggat cataagcgaa aggagaaatg
     2761 agatatgtct attcaaaaaa gtactaatag ttcaagttta gcagaagtca ttgaccgtat
     2821 tttagataaa ggaattgtta ttgatgcttt tgcaagagtt tctgttgtag gaattgaaat
     2881 tttaacgatt gaagcgcgag tggttattgc cagtgttgat acatggttac gctatgcaga
     2941 agcagtaggg cttcttcgtg acgacgtaga agaaaacggt cttcctgaac gttcaaattc
     3001 aagtgaaggg cagccgcgtt ttagtattta atatgaagtg gtatggaacc ttcctttttt
     3061 gagggtggtt ccagctttta gatctaacta ttggaggcta ctaaaaatgg aaattaaaaa
     3121 aattatgcaa gccgtgaacg actttttcgg tgaacacgta gctcctcctc ataaaattac
     3181 ctcggtggaa gctactgaag atgaaggttg gagagttatt gttgaagtca ttgaagaacg
     3241 agaatatatg aaaaaatacg ccaaagatga aatgctcgga acgtacgagt gctttgtaaa
     3301 taaagaaaaa gaagtcattt cattcaaacg actcgacgtc agatatagaa gcgccattgg
     3361 cattgaagca taaacgataa gatggcagga ggaacgtaaa aatgaccgtc ttaacagaca
     3421 aaaggaaaaa aggcagtgga gcttttatac aagatgacga gacaaaagag gttctttcaa
     3481 gagcgctgag ctatttaaaa tccggctatt ccattcattt tacaggtcct gccggcggag
     3541 gcaaaacctc tttagcgcga gcgcttgcta aaaagagaaa gcgtcctgta atgctgatgc
     3601 acgggaatca cgagctcaac aacaaagatt taattggcga ttttacggga tacacgagca
     3661 aaaaagtaat cgaccagtac gttcgttctg tctataaaaa agatgaacag gtgagtgaaa
     3721 actggcagga tggccgattg cttgaagctg taaaaaatgg ctatacgctg atttacgacg
     3781 aatttactcg ttctaagcct gcgacgaata atatctttct atcgatatta gaagaaggcg
     3841 tgctgccgct gtatggagta aaaatgaccg atccttttgt gcgcgtgcat cccgatttcc
     3901 gcgtcatctt cacaagcaat ccagctgagt atgccggcgt atatgatacg caagatgcgc
     3961 ttctcgacag gttaattacc atgtttattg attataaaga catcgacaga gagacagcga
     4021 ttttaacgga gaaaacggac gtagaagaag atgaagcgcg cacaattgta acgctcgtag
     4081 caaacgtgcg aaaccgctct ggagacgaaa acagcagcgg acttagcctg cgggcttcgc
     4141 ttatgatcgc tacccttgcc acgcagcaag acattcctat cgatggaagt gacgaagatt
     4201 ttcaaacgtt atgtatcgat attttgcatc atccgcttac caaatgtttg gatgaagaaa
     4261 atgcaaaaag caaagccgaa aaaatcattt tagaagaatg taagaatata gacactgaag
     4321 aaaagtaaag gagcttgaaa acatgagtga aacaaacgaa acaggtattt atatttttag
     4381 cgccattcaa acggataaag acgaagaatt tggcgccgtg gaagtagaag gaacaaaagc
     4441 tgaaacattt ttgattcgct acaaagacgc ggctatggta gcagctgaag taccgatgaa
     4501 aatttatcat cctaatcgcc aaaatttatt aatgcatcaa aacgcagtag cagcgattat
     4561 ggacaagaac gatacggtta ttccaatcag ctttgggaat gtattcaaat caaaagaaga
     4621 cgtaaaagtt cttttggaaa acctttatcc gcagtttgaa aagctgtttc cagcgatcaa
     4681 aggaaaaatt gaagtcggtt taaaagtaat tgggaaaaaa gaatggcttg agaaaaaagt
     4741 aaacgaaaat cctgaacttg agaaagtatc agcatccgta aaaggaaaat cagaagcagc
     4801 cggttattat gagcgtattc aacttggagg aatggctcaa aagatgttta cttccctgca
     4861 aaaagaagtc aagacagatg tgttttctcc gcttgaagaa gcagcggaag cagcaaaagc
     4921 aaatgagcca acgggcgaaa cgatgctttt aaacgcgtct ttcttaatta accgagaaga
     4981 tgaagcgaag tttgatgaaa aagtaaatga agcgcatgaa aactggaaag acaaagccga
     5041 ttttcattac agcggtcctt ggcctgctta taattttgtg aacattcgcc taaaagtaga
     5101 agagaaataa cgtgcttcac aaattagtaa ccgcacccat taaccttgta gtgaaaatcg
     5161 gcgaaaaagt acaggaagaa gctgataaac agctatatga ccttccgacg attcagcaaa
     5221 agctcattca gcttcaaatg atgtttgagc ttggtgaaat tccagaagaa gcgtttcaag
     5281 aaaaagaaga tgaattgtta atgaggtacg aaattgcgaa acgcagagaa attgaacaat
     5341 gggaagagct aacacaaaaa agaaatgagg aatcctagat gggagaatta ctgtatttat
     5401 acggtttaat tccaacaaaa gaagcagcag ccatagagcc gtttccatct tataaggggt
     5461 ttgacggaga acattcactg tacccaattg cgtttgatca ggtgacggct gtagtttcta
     5521 agctggatgc tgacacctat tcagaaaaag tgattcaaga aaaaatggag caggatatga
     5581 gctggctgca ggaaaaagca tttcatcatc acgaaacggt agccgctttg tacgaagaat
     5641 ttacgatcat tccattaaaa ttttgcacca tttataaagg tgaagaaagt ctgcaggcag
     5701 ctattgagat taacaaagaa aagatagaga attcactgac gctgcttcaa ggaaatgaag
     5761 agtggaatgt gaaaatttac tgtgatgata cagagcttaa aaaaggaatc agcgaaacga
     5821 atgaaagcgt gaaagcgaaa aaacaagaaa ttagtcactt atcaccagga agacagtttt
     5881 ttgaaaagaa aaaaatagat cagctgattg aaaaagaatt agagcttcac aaaaacaaag
     5941 tgtgtgaaga gatacatgac aagctaaaag aattatcgct ttatgactct gttaaaaaga
     6001 attggagcaa agacgtaact ggcgcagctg aacagatggc gtggaacagc gtgtttcttc
     6061 tcccgtctct gcagattact aagttcgtaa acgaaataga agagcttcag caaaggcttg
     6121 aaaataaagg ctggaagttt gaagtgacgg gaccatggcc gccctatcat ttctcgagct
     6181 ttgcgtaaag tgaggaatta acattatgtc tcttaaacaa tccatggaga ataaagatat
     6241 tgctcttatt gatattttag atgtcatttt agataaagga gtcgccatta aaggagactt
     6301 aatcatttcc atagctggcg tcgatttagt gtatttggat ttgcgggtgc ttatttcttc
     6361 ggttgaaacg cttgtgcaag caaaagaagg aaatcacaaa ccaatcactt ctgaacaatt
     6421 tgataaacaa aaggaggaat taatggatgc aaccggtcag ccaagcaaat ggacgaatcc
     6481 acttggatcc tgatcaagct gaacaaggct tagcgcagct tgtgatgaca gttattgagc
     6541 tattgaggca aatcgttgaa cgtcatgcca tgaggcgggt ggagggtgga acgttgacgg
     6601 acgaacaaat tgaaaactta ggaattgcac taatgaactt agaagaaaaa atggacgagt
     6661 tgaaagaggt gttcggtctg gatgcagaag atttaaatat tgatcttgga ccgctaggca
     6721 gcctgcttta agcggtcagt aggaggaaca gtatggcagt cgaacataat atgcagtcaa
     6781 gtacgattgt agatgtgctc gaaaagattt tggataaagg agtcgttata gcgggggaca
     6841 tcaccgtagg aattgcagat gtcgagctat taacgataaa gatccgcttg attgtggctt
     6901 cggttgataa ggcaaaagaa atcggcatgg actggtggga aaatgatccg tatctcagtt
     6961 caaaaggagc caataacaaa gcgctcgaag aagaaaataa aatgctgcat gagcggttaa
     7021 aaacgcttga agaaaaaata gaaacgaaac gttaaaaact gtacgctact taaaaaatgg
     7081 agggatttac aatggcaact gaaacaaaat tagataacac acaggcagaa aacaaggaaa
     7141 ataaaaatgc ggaaaacggt tcaaaagaaa agaacggttc aaaagcaagc aaaacaacaa
     7201 gcagcgggcc aatcaaacga gcggtagcag gaggcatcat cggtgcaacg attggatatg
     7261 tatcgactcc tgaaaatcga aaaagtctcc ttgaccgcat tgatacagac gaattaaaaa
     7321 gcaaagcatc tgatttagga acaaaggtaa aagaaaaatc aaaaagcagc gtggccagcc
     7381 tgaaaacatc tgcgggaagc ttgtttaaaa aagataaaga taaatcaaaa gatgatgaag
     7441 aaaacgtaaa ttcttctagt agcgaaacag aagacgataa cgttcaagag tacgacgagt
     7501 taaaagaaga aaatcaaact cttcaagatc gcttatcaca gcttgaagaa aaaatgaaca
     7561 tgcttgttga gcttagcctc aataaaaatc aagacgaaga agcggaagat acagattccg
     7621 acgaagaaga gaacgatgag aacgatgaaa acgatgaaaa cgagcaggac gatgaaaacg
     7681 aagaagaaac atctaagcca cgtaaaaagg ataaaaaaga agctgaggaa gaagaaagtg
     7741 aaagtgacga agacagcgag gaagaagagg aagattctcg ctcaaacaaa aaaaataaaa
     7801 aagtaaaaac agaagaagaa gacgaagatg aaagcgaaga agaaaaaaag gaagcgaaac
     7861 caaaaaagtc aacagctaaa aaatcaaaaa atacaaaagc aaagaaaaac acggacgaag
     7921 aagatgatga agcaacatct ctttctagtg aagacgatac aacagcctaa gacgtcgtca
     7981 catttgcttc tgcttgctgc agtccggatt cgcgaacgcg tctcgagaga tccggctgct
     8041 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     8101 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     8161 gactagtagc ccgcct
//
