LOCUS       sequence-445532-        8220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpB and GvpK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8220)
  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 8220)
  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'-deltaBK
FEATURES             Location/Qualifiers
     source          1..8220
                     /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(6478..6496)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6492..6539
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6606..6697
                     /gene="bla"
                     /label=AmpR promoter
     terminator      7562..7601
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      7632..8220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8121..8140
                     /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 taggtgctgg 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 aaaggaagag ttaatggacg cgaccggcca gccgagcaaa tggaccaatc
     4501 cgcttggctc ctgacccttg cgagcagtcg tatgctttct cgaattccga gtaggaggaa
     4561 cagtatggca gtcgaacata atatgcagtc aagtacgatt gtagatgtgc tcgaaaagat
     4621 tttggataaa ggagtcgtta tagcggggga catcaccgta ggaattgcag atgtcgagct
     4681 attaacgata aagatccgct tgattgtggc ttcggttgat aaggcaaaag aaatcggcat
     4741 ggactggtgg gaaaatgatc cgtatctcag ttcaaaagga gccaataaca aagcgctcga
     4801 agaagaaaat aaaatgctgc atgagcggtt aaaaacgctt gaagaaaaaa tagaaacgaa
     4861 acgttaaaaa ctgtacgcta cttaaaaaat ggagggattt acaatggcaa ctgaaacaaa
     4921 attagataac acacaggcag aaaacaagga aaataaaaat gcggaaaacg gttcaaaaga
     4981 aaagaacggt tcaaaagcaa gcaaaacaac aagcagcggg ccaatcaaac gagcggtagc
     5041 aggaggcatc atcggtgcaa cgattggata tgtatcgact cctgaaaatc gaaaaagtct
     5101 ccttgaccgc attgatacag acgaattaaa aagcaaagca tctgatttag gaacaaaggt
     5161 aaaagaaaaa tcaaaaagca gcgtggccag cctgaaaaca tctgcgggaa gcttgtttaa
     5221 aaaagataaa gataaatcaa aagatgatga agaaaacgta aattcttcta gtagcgaaac
     5281 agaagacgat aacgttcaag agtacgacga gttaaaagaa gaaaatcaaa ctcttcaaga
     5341 tcgcttatca cagcttgaag aaaaaatgaa catgcttgtt gagcttagcc tcaataaaaa
     5401 tcaagacgaa gaagcggaag atacagattc cgacgaagaa gagaacgatg agaacgatga
     5461 aaacgatgaa aacgagcagg acgatgaaaa cgaagaagaa acatctaagc cacgtaaaaa
     5521 ggataaaaaa gaagctgagg aagaagaaag tgaaagtgac gaagacagcg aggaagaaga
     5581 ggaagattct cgctcaaaca aaaaaaataa aaaagtaaaa acagaagaag aagacgaaga
     5641 tgaaagcgaa gaagaaaaaa aggaagcgaa accaaaaaag tcaacagcta aaaaatcaaa
     5701 aaatacaaaa gcaaagaaaa acacggacga agaagatgat gaagcaacat ctctttctag
     5761 tgaagacgat acaacagcct aagacgtaaa ggaggaaaga aagacatgag tacaggccct
     5821 tctttttcaa ctaaagacaa tacgcttgaa tactttgtga aagcttctaa taaacacggc
     5881 ttttcacttg atatttcatt aaatgtaaac ggcgctgtga tttccggtac catgatttca
     5941 gcaaaagaat actttgatta cttaagcgaa acgtttgaag aaggcagtga agtggctcag
     6001 gcgctaagcg aacaattctc tttagcaagc gaagcgagcg aatcaaacgg agaagcagaa
     6061 gcccatttta ttcatttgaa aaatacaaag atttactgtg gagacagtaa atctactcct
     6121 tctaaaggca aaatcttttg gagagggaaa atagcagaag tagacgggtt tttcttagga
     6181 aagatttctg atgcaaaatc aacgagtaaa aagagttcat aaaaaacggc ggggtgattg
     6241 ccccgccgtt ttttagtgat gtgatgagat gtgcagcttc ttttttcgat acatgagcgg
     6301 agtcatcttc atttcttttc gaaactgatt aataaaatag cttgtactat taaaccctac
     6361 ttgataggcg acttccgtca catttgcttc tgcttgctgc agtccggatt cgcgaacgcg
     6421 tctcgagaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     6481 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     6541 tgaaaggagg aactatatcc gactagtagc ccgcctaatg agcgggcttt tttttaattc
     6601 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6661 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6721 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6781 cgtgaaagta aaagacgcag aggaccaatt gggggcacga gtgggataca tagaactgga
     6841 cttgaatagc ggtaaaatcc ttgagagttt tcgccctgaa gagcgttttc caatgatgag
     6901 cactttcaaa gttctgctat gtggagcagt attatcccgt gtagatgcgg ggcaagagca
     6961 actcggacga cgaatacact attcgcagaa tgacttggtt gaatactccc cagtgacaga
     7021 aaagcacctt acggacggaa tgacggtaag agaattatgt agtgccgcca taacgatgag
     7081 tgataacact gcggcgaact tacttctgac aaccatcggt ggaccgaagg aattaaccgc
     7141 ttttttgcac aatatgggag accatgtaac tcgccttgac cgttgggaac cagaactgaa
     7201 tgaagccata ccaaacgacg agcgagacac cacaatgcct gcggcaatgg caacaacatt
     7261 acgcaaacta ttaactggcg aactacttac tctggcttca cggcaacaat taatagactg
     7321 gcttgaagcg gataaagttg caggaccact actgcgttcg gcacttcctg ctggctggtt
     7381 tattgctgat aaatctgggg caggagagcg tggttcacgg ggtatcattg ccgcacttgg
     7441 accagatggt aagccttccc gtatcgtagt tatctacacg acgggtagtc aggcaactat
     7501 ggacgaacga aatagacaga ttgctgaaat aggggcttca ctgattaagc attggtaaac
     7561 cgatacaatt aaaggctcct tttggagcct ttttttttgg acggaccggt agaaaagatc
     7621 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7681 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7741 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     7801 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7861 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7921 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7981 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8041 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8101 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8161 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
//
