LOCUS       sequence-446035-        7523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpB and GvpT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7523)
  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 7523)
  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'-deltaBT
FEATURES             Location/Qualifiers
     source          1..7523
                     /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             2927..2944
                     /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(5781..5799)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5795..5842
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5909..6000
                     /gene="bla"
                     /label=AmpR promoter
     terminator      6865..6904
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      6935..7523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7424..7443
                     /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 tgggatcatt agatctaact attggaggct actaaaaatg gaaattaaaa
     1141 aaattatgca agccgtgaac gactttttcg gtgaacacgt agctcctcct cataaaatta
     1201 cctcggtgga agctactgaa gatgaaggtt ggagagttat tgttgaagtc attgaagaac
     1261 gagaatatat gaaaaaatac gccaaagatg aaatgctcgg aacgtacgag tgctttgtaa
     1321 ataaagaaaa agaagtcatt tcattcaaac gactcgacgt cagatataga agcgccattg
     1381 gcattgaagc ataaacgata agatggcagg aggaacgtaa aaatgaccgt cttaacagac
     1441 aaaaggaaaa aaggcagtgg agcttttata caagatgacg agacaaaaga ggttctttca
     1501 agagcgctga gctatttaaa atccggctat tccattcatt ttacaggtcc tgccggcgga
     1561 ggcaaaacct ctttagcgcg agcgcttgct aaaaagagaa agcgtcctgt aatgctgatg
     1621 cacgggaatc acgagctcaa caacaaagat ttaattggcg attttacggg atacacgagc
     1681 aaaaaagtaa tcgaccagta cgttcgttct gtctataaaa aagatgaaca ggtgagtgaa
     1741 aactggcagg atggccgatt gcttgaagct gtaaaaaatg gctatacgct gatttacgac
     1801 gaatttactc gttctaagcc tgcgacgaat aatatctttc tatcgatatt agaagaaggc
     1861 gtgctgccgc tgtatggagt aaaaatgacc gatccttttg tgcgcgtgca tcccgatttc
     1921 cgcgtcatct tcacaagcaa tccagctgag tatgccggcg tatatgatac gcaagatgcg
     1981 cttctcgaca ggttaattac catgtttatt gattataaag acatcgacag agagacagcg
     2041 attttaacgg agaaaacgga cgtagaagaa gatgaagcgc gcacaattgt aacgctcgta
     2101 gcaaacgtgc gaaaccgctc tggagacgaa aacagcagcg gacttagcct gcgggcttcg
     2161 cttatgatcg ctacccttgc cacgcagcaa gacattccta tcgatggaag tgacgaagat
     2221 tttcaaacgt tatgtatcga tattttgcat catccgctta ccaaatgttt ggatgaagaa
     2281 aatgcaaaaa gcaaagccga aaaaatcatt ttagaagaat gtaagaatat agacactgaa
     2341 gaaaagtaaa ggagcttgaa aacatgagtg aaacaaacga aacaggtatt tatattttta
     2401 gcgccattca aacggataaa gacgaagaat ttggcgccgt ggaagtagaa ggaacaaaag
     2461 ctgaaacatt tttgattcgc tacaaagacg cggctatggt agcagctgaa gtaccgatga
     2521 aaatttatca tcctaatcgc caaaatttat taatgcatca aaacgcagta gcagcgatta
     2581 tggacaagaa cgatacggtt attccaatca gctttgggaa tgtattcaaa tcaaaagaag
     2641 acgtaaaagt tcttttggaa aacctttatc cgcagtttga aaagctgttt ccagcgatca
     2701 aaggaaaaat tgaagtcggt ttaaaagtaa ttgggaaaaa agaatggctt gagaaaaaag
     2761 taaacgaaaa tcctgaactt gagaaagtat cagcatccgt aaaaggaaaa tcagaagcag
     2821 ccggttatta tgagcgtatt caacttggag gaatggctca aaagatgttt acttccctgc
     2881 aaaaagaagt caagacagat gtgttttctc cgcttgaaga agcagcggaa gcagcaaaag
     2941 caaatgagcc aacgggcgaa acgatgcttt taaacgcgtc tttcttaatt aaccgagaag
     3001 atgaagcgaa gtttgatgaa aaagtaaatg aagcgcatga aaactggaaa gacaaagccg
     3061 attttcatta cagcggtcct tggcctgctt ataattttgt gaacattcgc ctaaaagtag
     3121 aagagaaata acgtgcttca caaattagta accgcaccca ttaaccttgt agtgaaaatc
     3181 ggcgaaaaag tacaggaaga agctgataaa cagctatatg accttccgac gattcagcaa
     3241 aagctcattc agcttcaaat gatgtttgag cttggtgaaa ttccagaaga agcgtttcaa
     3301 gaaaaagaag atgaattgtt aatgaggtac gaaattgcga aacgcagaga aattgaacaa
     3361 tgggaagagc taacacaaaa aagaaatgag gaatcctaga tgggagaatt actgtattta
     3421 tacggtttaa ttccaacaaa agaagcagca gccatagagc cgtttccatc ttataagggg
     3481 tttgacggag aacattcact gtacccaatt gcgtttgatc aggtgacggc tgtagtttct
     3541 aagctggatg ctgacaccta ttcagaaaaa gtgattcaag aaaaaatgga gcaggatatg
     3601 agctggctgc aggaaaaagc atttcatcat cacgaaacgg tagccgcttt gtacgaagaa
     3661 tttacgatca ttccattaaa attttgcacc atttataaag gtgaagaaag tctgcaggca
     3721 gctattgaga ttaacaaaga aaagatagag aattcactga cgctgcttca aggaaatgaa
     3781 gagtggaatg tgaaaattta ctgtgatgat acagagctta aaaaaggaat cagcgaaacg
     3841 aatgaaagcg tgaaagcgaa aaaacaagaa attagtcact tatcaccagg aagacagttt
     3901 tttgaaaaga aaaaaataga tcagctgatt gaaaaagaat tagagcttca caaaaacaaa
     3961 gtgtgtgaag agatacatga caagctaaaa gaattatcgc tttatgactc tgttaaaaag
     4021 aattggagca aagacgtaac tggcgcagct gaacagatgg cgtggaacag cgtgtttctt
     4081 ctcccgtctc tgcagattac taagttcgta aacgaaatag aagagcttca gcaaaggctt
     4141 gaaaataaag gctggaagtt tgaagtgacg ggaccatggc cgccctatca tttctcgagc
     4201 tttgcgtaaa gtgaggaatt aacattatgt ctcttaaaca atccatggag aataaagata
     4261 ttgctcttat tgatatttta gatgtcattt tagataaagg agtcgccatt aaaggagact
     4321 taatcatttc catagctggc gtcgatttag tgtatttgga tttgcgggtg cttatttctt
     4381 cggttgaaac gcttgtgcaa gcaaaagaag gaaatcacaa accaatcact tctgaacaat
     4441 ttgataaaca aaaggaggaa ttaatggatg caaccggtca gccaagcaaa tggacgaatc
     4501 cacttggatc ctgatcaagc tgaacaaggc ttagcgcagc ttgtgatgac agttattgag
     4561 ctattgaggc aaatcgttga acgtcatgcc atgaggcggg tggagggtgg aacgttgacg
     4621 gacgaacaaa ttgaaaactt aggaattgca ctaatgaact tagaagaaaa aatggacgag
     4681 ttgaaagagg tgttcggtct ggatgcagaa gatttaaata ttgatcttgg accgctaggc
     4741 agcctgcttt aagcggtcag taggaggaac agtatggcag tcgaacataa tatgcagtca
     4801 agtacgattg tagatgtgct cgaaaagatt ttggataaag gagtcgttat agcgggggac
     4861 atcaccgtag gaattgcaga tgtcgagcta ttaacgataa agatccgctt gattgtggct
     4921 tcggttgata aggcaaaaga aatcggcatg gactggtggg aaaatgatcc gtatctcagt
     4981 tcaaaaggag ccaataacaa agcgctcgaa gaagaaaata aaatgctgca tgagcggtta
     5041 aaaacgcttg aagaaaaaat agaaacgaaa cgttaaaaac tgtacgctac ttaggaggaa
     5101 agaaagacat gagtacaggc ccttcttttt caactaaaga caatacgctt gaatactttg
     5161 tgaaagcttc taataaacac ggcttttcac ttgatatttc attaaatgta aacggcgctg
     5221 tgatttccgg taccatgatt tcagcaaaag aatactttga ttacttaagc gaaacgtttg
     5281 aagaaggcag tgaagtggct caggcgctaa gcgaacaatt ctctttagca agcgaagcga
     5341 gcgaatcaaa cggagaagca gaagcccatt ttattcattt gaaaaataca aagatttact
     5401 gtggagacag taaatctact ccttctaaag gcaaaatctt ttggagaggg aaaatagcag
     5461 aagtagacgg gtttttctta ggaaagattt ctgatgcaaa atcaacgagt aaaaagagtt
     5521 cataaaaaac ggcggggtga ttgccccgcc gttttttagt gatgtgatga gatgtgcagc
     5581 ttcttttttc gatacatgag cggagtcatc ttcatttctt ttcgaaactg attaataaaa
     5641 tagcttgtac tattaaaccc tacttgatag gcgacttccg tcacatttgc ttctgcttgc
     5701 tgcagtccgg attcgcgaac gcgtctcgag agatccggct gctaacaaag cccgaaagga
     5761 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     5821 acgggtcttg aggggttttt tgctgaaagg aggaactata tccgactagt agcccgccta
     5881 atgagcgggc ttttttttaa ttcccctatt tgtttatttt tctaaataca ttcaaatatg
     5941 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6001 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6061 gtttttgctc acccagaaac gctcgtgaaa gtaaaagacg cagaggacca attgggggca
     6121 cgagtgggat acatagaact ggacttgaat agcggtaaaa tccttgagag ttttcgccct
     6181 gaagagcgtt ttccaatgat gagcactttc aaagttctgc tatgtggagc agtattatcc
     6241 cgtgtagatg cggggcaaga gcaactcgga cgacgaatac actattcgca gaatgacttg
     6301 gttgaatact ccccagtgac agaaaagcac cttacggacg gaatgacggt aagagaatta
     6361 tgtagtgccg ccataacgat gagtgataac actgcggcga acttacttct gacaaccatc
     6421 ggtggaccga aggaattaac cgcttttttg cacaatatgg gagaccatgt aactcgcctt
     6481 gaccgttggg aaccagaact gaatgaagcc ataccaaacg acgagcgaga caccacaatg
     6541 cctgcggcaa tggcaacaac attacgcaaa ctattaactg gcgaactact tactctggct
     6601 tcacggcaac aattaataga ctggcttgaa gcggataaag ttgcaggacc actactgcgt
     6661 tcggcacttc ctgctggctg gtttattgct gataaatctg gggcaggaga gcgtggttca
     6721 cggggtatca ttgccgcact tggaccagat ggtaagcctt cccgtatcgt agttatctac
     6781 acgacgggta gtcaggcaac tatggacgaa cgaaatagac agattgctga aataggggct
     6841 tcactgatta agcattggta aaccgataca attaaaggct ccttttggag cctttttttt
     6901 tggacggacc ggtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6961 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7021 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7081 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7141 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7201 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7261 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7321 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7381 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7441 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7501 caggggggcg gagcctatgg aaa
//
