LOCUS       sequence-445900-        8563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpJ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8563)
  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 8563)
  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'-deltaJ
FEATURES             Location/Qualifiers
     source          1..8563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             35..226
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(220..242)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    328..467
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     383..402
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(482..499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(653..1241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(733..752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1272..1311)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(2176..2267)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2334..2381)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2377..2395
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5937..5954)
                     /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(8342..8361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8343..8361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct
       61 taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct
      121 ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta
      181 ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      241 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      301 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
      361 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc
      421 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
      481 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      541 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      601 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      661 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      721 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      781 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      841 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      901 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      961 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1021 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1081 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1141 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1201 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1261 ctaccggtcc gtccaaaaaa aaaggctcca aaaggagcct ttaattgtat cggtttacca
     1321 atgcttaatc agtgaagccc ctatttcagc aatctgtcta tttcgttcgt ccatagttgc
     1381 ctgactaccc gtcgtgtaga taactacgat acgggaaggc ttaccatctg gtccaagtgc
     1441 ggcaatgata ccccgtgaac cacgctctcc tgccccagat ttatcagcaa taaaccagcc
     1501 agcaggaagt gccgaacgca gtagtggtcc tgcaacttta tccgcttcaa gccagtctat
     1561 taattgttgc cgtgaagcca gagtaagtag ttcgccagtt aatagtttgc gtaatgttgt
     1621 tgccattgcc gcaggcattg tggtgtctcg ctcgtcgttt ggtatggctt cattcagttc
     1681 tggttcccaa cggtcaaggc gagttacatg gtctcccata ttgtgcaaaa aagcggttaa
     1741 ttccttcggt ccaccgatgg ttgtcagaag taagttcgcc gcagtgttat cactcatcgt
     1801 tatggcggca ctacataatt ctcttaccgt cattccgtcc gtaaggtgct tttctgtcac
     1861 tggggagtat tcaaccaagt cattctgcga atagtgtatt cgtcgtccga gttgctcttg
     1921 ccccgcatct acacgggata atactgctcc acatagcaga actttgaaag tgctcatcat
     1981 tggaaaacgc tcttcagggc gaaaactctc aaggatttta ccgctattca agtccagttc
     2041 tatgtatccc actcgtgccc ccaattggtc ctctgcgtct tttactttca cgagcgtttc
     2101 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2161 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2221 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggaattaaa
     2281 aaaaagcccg ctcattaggc gggctactag tcggatatag ttcctccttt cagcaaaaaa
     2341 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca
     2401 gcagccaact cagcttcctt tcgggctttg ttagcagccg gatctctcga gacgcgttcg
     2461 cgaatccgga ctgcagcaag cagaagcaaa tgtgacggaa gtcgcctatc aagtagggtt
     2521 taatagtaca agctatttta ttaatcagtt tcgaaaagaa atgaagatga ctccgctcat
     2581 gtatcgaaaa aagaagctgc acatctcatc acatcactaa aaaacggcgg ggcaatcacc
     2641 ccgccgtttt ttatgaactc tttttactcg ttgattttgc atcagaaatc tttcctaaga
     2701 aaaacccgtc tacttctgct attttccctc tccaaaagat tttgccttta gaaggagtag
     2761 atttactgtc tccacagtaa atctttgtat ttttcaaatg aataaaatgg gcttctgctt
     2821 ctccgtttga ttcgctcgct tcgcttgcta aagagaattg ttcgcttagc gcctgagcca
     2881 cttcactgcc ttcttcaaac gtttcgctta agtaatcaaa gtattctttt gctgaaatca
     2941 tggtaccgga aatcacagcg ccgtttacat ttaatgaaat atcaagtgaa aagccgtgtt
     3001 tattagaagc tttcacaaag tattcaagcg tattgtcttt agttgaaaaa gaagggcctg
     3061 tactcatgtc tttctttcct cctttacgtc ttaggctgtt gtatcgtctt cactagaaag
     3121 agatgttgct tcatcatctt cttcgtccgt gtttttcttt gcttttgtat tttttgattt
     3181 tttagctgtt gacttttttg gtttcgcttc ctttttttct tcttcgcttt catcttcgtc
     3241 ttcttcttct gtttttactt ttttattttt tttgtttgag cgagaatctt cctcttcttc
     3301 ctcgctgtct tcgtcacttt cactttcttc ttcctcagct tcttttttat cctttttacg
     3361 tggcttagat gtttcttctt cgttttcatc gtcctgctcg ttttcatcgt tttcatcgtt
     3421 ctcatcgttc tcttcttcgt cggaatctgt atcttccgct tcttcgtctt gatttttatt
     3481 gaggctaagc tcaacaagca tgttcatttt ttcttcaagc tgtgataagc gatcttgaag
     3541 agtttgattt tcttctttta actcgtcgta ctcttgaacg ttatcgtctt ctgtttcgct
     3601 actagaagaa tttacgtttt cttcatcatc ttttgattta tctttatctt ttttaaacaa
     3661 gcttcccgca gatgttttca ggctggccac gctgcttttt gatttttctt ttacctttgt
     3721 tcctaaatca gatgctttgc tttttaattc gtctgtatca atgcggtcaa ggagactttt
     3781 tcgattttca ggagtcgata catatccaat cgttgcaccg atgatgcctc ctgctaccgc
     3841 tcgtttgatt ggcccgctgc ttgttgtttt gcttgctttt gaaccgttct tttcttttga
     3901 accgttttcc gcatttttat tttccttgtt ttctgcctgt gtgttatcta attttgtttc
     3961 agttgccatt gtaaatccct ccatttttta agtagcgtac agctgaccgc ttaaagcagg
     4021 ctgcctagcg ggccaagatc aatatttaaa tcttctgcat ccagaccgaa cacctctttc
     4081 aactcgtcca ttttttcttc taagttcatt agtgcaattc ctaagttttc aatttgttcg
     4141 tccgtcaacg ttccaccctc cacccgcctc atggcatgac gttcaacgat ttgcctcaat
     4201 agctcaataa ctgtcatcac aagctgcgct aagccttgtt cagcttgatc aggatccaag
     4261 tggattcgtc catttgcttg gctgaccggt tgcatctatt aattcctcct ggactgtcac
     4321 gcttaaccgc tcggaattcg agaaagcata cgactgctcg caagggtcac gaacctagag
     4381 ggttcgtcca cttactaggc tggcccgtag cgtccatcag ctcttctttt tgtttatcaa
     4441 attgttcaga agtgattggt ttgtgatttc cttcttttgc ttgcacaagc gtttcaaccg
     4501 aagaaataag cacccgcaaa tccaaataca ctaaatcgac gccagctatg gaaatgatta
     4561 agtctccttt aatggcgact cctttatcta aaatgacatc taaaatatca ataagagcaa
     4621 tatctttatt ctccatggat tgtttaagag acataatgtt aattcctcac tttacgcaaa
     4681 gctcgagaaa tgatagggcg gccatggtcc cgtcacttca aacttccagc ctttattttc
     4741 aagcctttgc tgaagctctt ctatttcgtt tacgaactta gtaatctgca gagacgggag
     4801 aagaaacacg ctgttccacg ccatctgttc agctgcgcca gttacgtctt tgctccaatt
     4861 ctttttaaca gagtcataaa gcgataattc ttttagcttg tcatgtatct cttcacacac
     4921 tttgtttttg tgaagctcta attctttttc aatcagctga tctatttttt tcttttcaaa
     4981 aaactgtctt cctggtgata agtgactaat ttcttgtttt ttcgctttca cgctttcatt
     5041 cgtttcgctg attccttttt taagctctgt atcatcacag taaattttca cattccactc
     5101 ttcatttcct tgaagcagcg tcagtgaatt ctctatcttt tctttgttaa tctcaatagc
     5161 tgcctgcaga ctttcttcac ctttataaat ggtgcaaaat tttaatggaa tgatcgtaaa
     5221 ttcttcgtac aaagcggcta ccgtttcgtg atgatgaaat gctttttcct gcagccagct
     5281 catatcctgc tccatttttt cttgaatcac tttttctgaa taggtgtcag catccagctt
     5341 agaaactaca gccgtcacct gatcaaacgc aattgggtac agtgaatgtt ctccgtcaaa
     5401 ccccttataa gatggaaacg gctctatggc tgctgcttct tttgttggaa ttaaaccgta
     5461 taaatacagt aattctccca tctaggattc ctcatttctt ttttgtgtta gctcttccca
     5521 ttgttcaatt tctctgcgtt tcgcaatttc gtacctcatt aacaattcat cttctttttc
     5581 ttgaaacgct tcttctggaa tttcaccaag ctcaaacatc atttgaagct gaatgagctt
     5641 ttgctgaatc gtcggaaggt catatagctg tttatcagct tcttcctgta ctttttcgcc
     5701 gattttcact acaaggttaa tgggtgcggt tactaatttg tgaagcacgt tatttctctt
     5761 ctacttttag gcgaatgttc acaaaattat aagcaggcca aggaccgctg taatgaaaat
     5821 cggctttgtc tttccagttt tcatgcgctt catttacttt ttcatcaaac ttcgcttcat
     5881 cttctcggtt aattaagaaa gacgcgttta aaagcatcgt ttcgcccgtt ggctcatttg
     5941 cttttgctgc ttccgctgct tcttcaagcg gagaaaacac atctgtcttg acttcttttt
     6001 gcagggaagt aaacatcttt tgagccattc ctccaagttg aatacgctca taataaccgg
     6061 ctgcttctga ttttcctttt acggatgctg atactttctc aagttcagga ttttcgttta
     6121 cttttttctc aagccattct tttttcccaa ttacttttaa accgacttca atttttcctt
     6181 tgatcgctgg aaacagcttt tcaaactgcg gataaaggtt ttccaaaaga acttttacgt
     6241 cttcttttga tttgaataca ttcccaaagc tgattggaat aaccgtatcg ttcttgtcca
     6301 taatcgctgc tactgcgttt tgatgcatta ataaattttg gcgattagga tgataaattt
     6361 tcatcggtac ttcagctgct accatagccg cgtctttgta gcgaatcaaa aatgtttcag
     6421 cttttgttcc ttctacttcc acggcgccaa attcttcgtc tttatccgtt tgaatggcgc
     6481 taaaaatata aatacctgtt tcgtttgttt cactcatgtt ttcaagctcc tttacttttc
     6541 ttcagtgtct atattcttac attcttctaa aatgattttt tcggctttgc tttttgcatt
     6601 ttcttcatcc aaacatttgg taagcggatg atgcaaaata tcgatacata acgtttgaaa
     6661 atcttcgtca cttccatcga taggaatgtc ttgctgcgtg gcaagggtag cgatcataag
     6721 cgaagcccgc aggctaagtc cgctgctgtt ttcgtctcca gagcggtttc gcacgtttgc
     6781 tacgagcgtt acaattgtgc gcgcttcatc ttcttctacg tccgttttct ccgttaaaat
     6841 cgctgtctct ctgtcgatgt ctttataatc aataaacatg gtaattaacc tgtcgagaag
     6901 cgcatcttgc gtatcatata cgccggcata ctcagctgga ttgcttgtga agatgacgcg
     6961 gaaatcggga tgcacgcgca caaaaggatc ggtcattttt actccataca gcggcagcac
     7021 gccttcttct aatatcgata gaaagatatt attcgtcgca ggcttagaac gagtaaattc
     7081 gtcgtaaatc agcgtatagc cattttttac agcttcaagc aatcggccat cctgccagtt
     7141 ttcactcacc tgttcatctt ttttatagac agaacgaacg tactggtcga ttactttttt
     7201 gctcgtgtat cccgtaaaat cgccaattaa atctttgttg ttgagctcgt gattcccgtg
     7261 catcagcatt acaggacgct ttctcttttt agcaagcgct cgcgctaaag aggttttgcc
     7321 tccgccggca ggacctgtaa aatgaatgga atagccggat tttaaatagc tcagcgctct
     7381 tgaaagaacc tcttttgtct cgtcatcttg tataaaagct ccactgcctt ttttcctttt
     7441 gtctgttaag acggtcattt ttacgttcct cctgccatct tatcgtttat gcttcaatgc
     7501 caatggcgct tctatatctg acgtcgagtc gtttgaatga aatgacttct ttttctttat
     7561 ttacaaagca ctcgtacgtt ccgagcattt catctttggc gtattttttc atatattctc
     7621 gttcttcaat gacttcaaca ataactctcc aaccttcatc ttcagtagct tccaccgagg
     7681 taattttatg aggaggagct acgtgttcac cgaaaaagtc gttcacggct tgcataattt
     7741 ttttaatttc catttttagt agcctccaat agttagatct aaaagctgga accaccctca
     7801 aaaaaggaag gttccatacc acttcatatt aaatactaaa acgcggctgc ccttcacttg
     7861 aatttgaacg ttcaggaaga ccgttttctt ctacgtcgtc acgaagaagc cctactgctt
     7921 ctgcatagcg taaccatgta tcaacactgg caataaccac tcgcgcttca atcgttaaaa
     7981 tttcaattcc tacaacagaa actcttgcaa aagcatcaat aacaattcct ttatctaaaa
     8041 tacggtcaat gacttctgct aaacttgaac tattagtact tttttgaata gacatatctc
     8101 atttctcctt tcgcttatga tcccattgtt tttaaatctg tagatgattt aatatcatta
     8161 gaacttttaa tcttagtaga gcttttaata tccgttgctt ttttaatctg atttacgcct
     8221 tttattttcc gcttattttt tgacttctct ttatcgtctg aagaagaaac tttttcttga
     8281 aagttttctc cagcttcttt cacatcgcta agctatctct agagggaaac cgttgtggtc
     8341 tccctatagt gagtcgtatt aatttcgcgg gatcgatgca tggtgccaac acctacatct
     8401 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     8461 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     8521 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta ccc
//
