LOCUS       sequence-446040-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpR and GvpL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7647)
  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 7647)
  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'-deltaRL
FEATURES             Location/Qualifiers
     source          1..7647
                     /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(5905..5923)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5919..5966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6033..6124
                     /gene="bla"
                     /label=AmpR promoter
     terminator      6989..7028
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      7059..7647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7548..7567
                     /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 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 atcctagagt gaggaattaa cattatgtct cttaaacaat ccatggagaa taaagatatt
     3481 gctcttattg atattttaga tgtcatttta gataaaggag tcgccattaa aggagactta
     3541 atcatttcca tagctggcgt cgatttagtg tatttggatt tgcgggtgct tatttcttcg
     3601 gttgaaacgc ttgtgcaagc aaaagaagga aatcacaaac caatcacttc tgaacaattt
     3661 gataaacaaa aggaggaatt aatggatgca accggtcagc caagcaaatg gacgaatcca
     3721 cttggatcct gatcaagctg aacaaggctt agcgcagctt gtgatgacag ttattgagct
     3781 attgaggcaa atcgttgaac gtcatgccat gaggcgggtg gagggtggaa cgttgacgga
     3841 cgaacaaatt gaaaacttag gaattgcact aatgaactta gaagaaaaaa tggacgagtt
     3901 gaaagaggtg ttcggtctgg atgcagaaga tttaaatatt gatcttggac cgctaggcag
     3961 cctgctttaa gcggtcagta ggaggaacag tatggcagtc gaacataata tgcagtcaag
     4021 tacgattgta gatgtgctcg aaaagatttt ggataaagga gtcgttatag cgggggacat
     4081 caccgtagga attgcagatg tcgagctatt aacgataaag atccgcttga ttgtggcttc
     4141 ggttgataag gcaaaagaaa tcggcatgga ctggtgggaa aatgatccgt atctcagttc
     4201 aaaaggagcc aataacaaag cgctcgaaga agaaaataaa atgctgcatg agcggttaaa
     4261 aacgcttgaa gaaaaaatag aaacgaaacg ttaaaaactg tacgctactt aaaaaatgga
     4321 gggatttaca atggcaactg aaacaaaatt agataacaca caggcagaaa acaaggaaaa
     4381 taaaaatgcg gaaaacggtt caaaagaaaa gaacggttca aaagcaagca aaacaacaag
     4441 cagcgggcca atcaaacgag cggtagcagg aggcatcatc ggtgcaacga ttggatatgt
     4501 atcgactcct gaaaatcgaa aaagtctcct tgaccgcatt gatacagacg aattaaaaag
     4561 caaagcatct gatttaggaa caaaggtaaa agaaaaatca aaaagcagcg tggccagcct
     4621 gaaaacatct gcgggaagct tgtttaaaaa agataaagat aaatcaaaag atgatgaaga
     4681 aaacgtaaat tcttctagta gcgaaacaga agacgataac gttcaagagt acgacgagtt
     4741 aaaagaagaa aatcaaactc ttcaagatcg cttatcacag cttgaagaaa aaatgaacat
     4801 gcttgttgag cttagcctca ataaaaatca agacgaagaa gcggaagata cagattccga
     4861 cgaagaagag aacgatgaga acgatgaaaa cgatgaaaac gagcaggacg atgaaaacga
     4921 agaagaaaca tctaagccac gtaaaaagga taaaaaagaa gctgaggaag aagaaagtga
     4981 aagtgacgaa gacagcgagg aagaagagga agattctcgc tcaaacaaaa aaaataaaaa
     5041 agtaaaaaca gaagaagaag acgaagatga aagcgaagaa gaaaaaaagg aagcgaaacc
     5101 aaaaaagtca acagctaaaa aatcaaaaaa tacaaaagca aagaaaaaca cggacgaaga
     5161 agatgatgaa gcaacatctc tttctagtga agacgataca acagcctaag acgtaaagga
     5221 ggaaagaaag acatgagtac aggcccttct ttttcaacta aagacaatac gcttgaatac
     5281 tttgtgaaag cttctaataa acacggcttt tcacttgata tttcattaaa tgtaaacggc
     5341 gctgtgattt ccggtaccat gatttcagca aaagaatact ttgattactt aagcgaaacg
     5401 tttgaagaag gcagtgaagt ggctcaggcg ctaagcgaac aattctcttt agcaagcgaa
     5461 gcgagcgaat caaacggaga agcagaagcc cattttattc atttgaaaaa tacaaagatt
     5521 tactgtggag acagtaaatc tactccttct aaaggcaaaa tcttttggag agggaaaata
     5581 gcagaagtag acgggttttt cttaggaaag atttctgatg caaaatcaac gagtaaaaag
     5641 agttcataaa aaacggcggg gtgattgccc cgccgttttt tagtgatgtg atgagatgtg
     5701 cagcttcttt tttcgataca tgagcggagt catcttcatt tcttttcgaa actgattaat
     5761 aaaatagctt gtactattaa accctacttg ataggcgact tccgtcacat ttgcttctgc
     5821 ttgctgcagt ccggattcgc gaacgcgtct cgagagatcc ggctgctaac aaagcccgaa
     5881 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     5941 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgac tagtagcccg
     6001 cctaatgagc gggctttttt ttaattcccc tatttgttta tttttctaaa tacattcaaa
     6061 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6121 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6181 tcctgttttt gctcacccag aaacgctcgt gaaagtaaaa gacgcagagg accaattggg
     6241 ggcacgagtg ggatacatag aactggactt gaatagcggt aaaatccttg agagttttcg
     6301 ccctgaagag cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gagcagtatt
     6361 atcccgtgta gatgcggggc aagagcaact cggacgacga atacactatt cgcagaatga
     6421 cttggttgaa tactccccag tgacagaaaa gcaccttacg gacggaatga cggtaagaga
     6481 attatgtagt gccgccataa cgatgagtga taacactgcg gcgaacttac ttctgacaac
     6541 catcggtgga ccgaaggaat taaccgcttt tttgcacaat atgggagacc atgtaactcg
     6601 ccttgaccgt tgggaaccag aactgaatga agccatacca aacgacgagc gagacaccac
     6661 aatgcctgcg gcaatggcaa caacattacg caaactatta actggcgaac tacttactct
     6721 ggcttcacgg caacaattaa tagactggct tgaagcggat aaagttgcag gaccactact
     6781 gcgttcggca cttcctgctg gctggtttat tgctgataaa tctggggcag gagagcgtgg
     6841 ttcacggggt atcattgccg cacttggacc agatggtaag ccttcccgta tcgtagttat
     6901 ctacacgacg ggtagtcagg caactatgga cgaacgaaat agacagattg ctgaaatagg
     6961 ggcttcactg attaagcatt ggtaaaccga tacaattaaa ggctcctttt ggagcctttt
     7021 tttttggacg gaccggtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7081 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7141 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7201 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7261 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7321 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7381 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7441 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7501 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7561 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7621 tcgtcagggg ggcggagcct atggaaa
//
