LOCUS       sequence-445450-        7553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpR and GvpT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7553)
  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 7553)
  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'-deltaRT
FEATURES             Location/Qualifiers
     source          1..7553
                     /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(5811..5829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5825..5872
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5939..6030
                     /gene="bla"
                     /label=AmpR promoter
     terminator      6895..6934
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      6965..7553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7454..7473
                     /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 aagtcattta 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 tgcgtaaagt gaggaattaa cattatgtct
     4261 cttaaacaat ccatggagaa taaagatatt gctcttattg atattttaga tgtcatttta
     4321 gataaaggag tcgccattaa aggagactta atcatttcca tagctggcgt cgatttagtg
     4381 tatttggatt tgcgggtgct tatttcttcg gttgaaacgc ttgtgcaagc aaaagaagga
     4441 aatcacaaac caatcacttc tgaacaattt gataaacaaa aggaggaatt aatggatgca
     4501 accggtcagc caagcaaatg gacgaatcca cttggatcct gatcaagctg aacaaggctt
     4561 agcgcagctt gtgatgacag ttattgagct attgaggcaa atcgttgaac gtcatgccat
     4621 gaggcgggtg gagggtggaa cgttgacgga cgaacaaatt gaaaacttag gaattgcact
     4681 aatgaactta gaagaaaaaa tggacgagtt gaaagaggtg ttcggtctgg atgcagaaga
     4741 tttaaatatt gatcttggac cgctaggcag cctgctttaa gcggtcagta ggaggaacag
     4801 tatggcagtc gaacataata tgcagtcaag tacgattgta gatgtgctcg aaaagatttt
     4861 ggataaagga gtcgttatag cgggggacat caccgtagga attgcagatg tcgagctatt
     4921 aacgataaag atccgcttga ttgtggcttc ggttgataag gcaaaagaaa tcggcatgga
     4981 ctggtgggaa aatgatccgt atctcagttc aaaaggagcc aataacaaag cgctcgaaga
     5041 agaaaataaa atgctgcatg agcggttaaa aacgcttgaa gaaaaaatag aaacgaaacg
     5101 ttaaaaactg tacgctactt aaaggaggaa agaaagacat gagtacaggc ccttcttttt
     5161 caactaaaga caatacgctt gaatactttg tgaaagcttc taataaacac ggcttttcac
     5221 ttgatatttc attaaatgta aacggcgctg tgatttccgg taccatgatt tcagcaaaag
     5281 aatactttga ttacttaagc gaaacgtttg aagaaggcag tgaagtggct caggcgctaa
     5341 gcgaacaatt ctctttagca agcgaagcga gcgaatcaaa cggagaagca gaagcccatt
     5401 ttattcattt gaaaaataca aagatttact gtggagacag taaatctact ccttctaaag
     5461 gcaaaatctt ttggagaggg aaaatagcag aagtagacgg gtttttctta ggaaagattt
     5521 ctgatgcaaa atcaacgagt aaaaagagtt cataaaaaac ggcggggtga ttgccccgcc
     5581 gttttttagt gatgtgatga gatgtgcagc ttcttttttc gatacatgag cggagtcatc
     5641 ttcatttctt ttcgaaactg attaataaaa tagcttgtac tattaaaccc tacttgatag
     5701 gcgacttccg tcacatttgc ttctgcttgc tgcagtccgg attcgcgaac gcgtctcgag
     5761 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     5821 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     5881 aggaactata tccgactagt agcccgccta atgagcgggc ttttttttaa ttcccctatt
     5941 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6001 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6061 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctcgtgaaa
     6121 gtaaaagacg cagaggacca attgggggca cgagtgggat acatagaact ggacttgaat
     6181 agcggtaaaa tccttgagag ttttcgccct gaagagcgtt ttccaatgat gagcactttc
     6241 aaagttctgc tatgtggagc agtattatcc cgtgtagatg cggggcaaga gcaactcgga
     6301 cgacgaatac actattcgca gaatgacttg gttgaatact ccccagtgac agaaaagcac
     6361 cttacggacg gaatgacggt aagagaatta tgtagtgccg ccataacgat gagtgataac
     6421 actgcggcga acttacttct gacaaccatc ggtggaccga aggaattaac cgcttttttg
     6481 cacaatatgg gagaccatgt aactcgcctt gaccgttggg aaccagaact gaatgaagcc
     6541 ataccaaacg acgagcgaga caccacaatg cctgcggcaa tggcaacaac attacgcaaa
     6601 ctattaactg gcgaactact tactctggct tcacggcaac aattaataga ctggcttgaa
     6661 gcggataaag ttgcaggacc actactgcgt tcggcacttc ctgctggctg gtttattgct
     6721 gataaatctg gggcaggaga gcgtggttca cggggtatca ttgccgcact tggaccagat
     6781 ggtaagcctt cccgtatcgt agttatctac acgacgggta gtcaggcaac tatggacgaa
     6841 cgaaatagac agattgctga aataggggct tcactgatta agcattggta aaccgataca
     6901 attaaaggct ccttttggag cctttttttt tggacggacc ggtagaaaag atcaaaggat
     6961 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7021 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7081 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     7141 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7201 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7261 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7321 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7381 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7441 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7501 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaa
//
