LOCUS       sequence-445901-        8457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mega' with knockout of GvpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8457)
  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 8457)
  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'-deltaR
FEATURES             Location/Qualifiers
     source          1..8457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             713..730
                     /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(4473..4491)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4487..4534
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        4601..4692
                     /gene="bla"
                     /label=AmpR promoter
     terminator      5557..5596
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      5627..6215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6116..6135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6369..6386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6401..6540
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6466..6485)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6626..6648
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6642..6833)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     7070..7089
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7070..7088
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaagattttc aaacgttatg tatcgatatt ttgcatcatc cgcttaccaa atgtttggat
       61 gaagaaaatg caaaaagcaa agccgaaaaa atcattttag aagaatgtaa gaatatagac
      121 actgaagaaa agtaaaggag cttgaaaaca tgagtgaaac aaacgaaaca ggtatttata
      181 tttttagcgc cattcaaacg gataaagacg aagaatttgg cgccgtggaa gtagaaggaa
      241 caaaagctga aacatttttg attcgctaca aagacgcggc tatggtagca gctgaagtac
      301 cgatgaaaat ttatcatcct aatcgccaaa atttattaat gcatcaaaac gcagtagcag
      361 cgattatgga caagaacgat acggttattc caatcagctt tgggaatgta ttcaaatcaa
      421 aagaagacgt aaaagttctt ttggaaaacc tttatccgca gtttgaaaag ctgtttccag
      481 cgatcaaagg aaaaattgaa gtcggtttaa aagtaattgg gaaaaaagaa tggcttgaga
      541 aaaaagtaaa cgaaaatcct gaacttgaga aagtatcagc atccgtaaaa ggaaaatcag
      601 aagcagccgg ttattatgag cgtattcaac ttggaggaat ggctcaaaag atgtttactt
      661 ccctgcaaaa agaagtcaag acagatgtgt tttctccgct tgaagaagca gcggaagcag
      721 caaaagcaaa tgagccaacg ggcgaaacga tgcttttaaa cgcgtctttc ttaattaacc
      781 gagaagatga agcgaagttt gatgaaaaag taaatgaagc gcatgaaaac tggaaagaca
      841 aagccgattt tcattacagc ggtccttggc ctgcttataa ttttgtgaac attcgcctaa
      901 aagtagaaga gaaataacgt gcttcacaaa ttagtaaccg cacccattaa ccttgtagtg
      961 aaaatcggcg aaaaagtaca ggaagaagct gataaacagc tatatgacct tccgacgatt
     1021 cagcaaaagc tcattcagct tcaaatgatg tttgagcttg gtgaaattcc agaagaagcg
     1081 tttcaagaaa aagaagatga attgttaatg aggtacgaaa ttgcgaaacg cagagaaatt
     1141 gaacaatggg aagagctaac acaaaaaaga aatgaggaat cctagatggg agaattactg
     1201 tatttatacg gtttaattcc aacaaaagaa gcagcagcca tagagccgtt tccatcttat
     1261 aaggggtttg acggagaaca ttcactgtac ccaattgcgt ttgatcaggt gacggctgta
     1321 gtttctaagc tggatgctga cacctattca gaaaaagtga ttcaagaaaa aatggagcag
     1381 gatatgagct ggctgcagga aaaagcattt catcatcacg aaacggtagc cgctttgtac
     1441 gaagaattta cgatcattcc attaaaattt tgcaccattt ataaaggtga agaaagtctg
     1501 caggcagcta ttgagattaa caaagaaaag atagagaatt cactgacgct gcttcaagga
     1561 aatgaagagt ggaatgtgaa aatttactgt gatgatacag agcttaaaaa aggaatcagc
     1621 gaaacgaatg aaagcgtgaa agcgaaaaaa caagaaatta gtcacttatc accaggaaga
     1681 cagttttttg aaaagaaaaa aatagatcag ctgattgaaa aagaattaga gcttcacaaa
     1741 aacaaagtgt gtgaagagat acatgacaag ctaaaagaat tatcgcttta tgactctgtt
     1801 aaaaagaatt ggagcaaaga cgtaactggc gcagctgaac agatggcgtg gaacagcgtg
     1861 tttcttctcc cgtctctgca gattactaag ttcgtaaacg aaatagaaga gcttcagcaa
     1921 aggcttgaaa ataaaggctg gaagtttgaa gtgacgggac catggccgcc ctatcatttc
     1981 tcgagctttg cgtaaagtga ggaattaaca ttatgtctct taaacaatcc atggagaata
     2041 aagatattgc tcttattgat attttagatg tcattttaga taaaggagtc gccattaaag
     2101 gagacttaat catttccata gctggcgtcg atttagtgta tttggatttg cgggtgctta
     2161 tttcttcggt tgaaacgctt gtgcaagcaa aagaaggaaa tcacaaacca atcacttctg
     2221 aacaatttga taaacaaaag gaggaattaa tggatgcaac cggtcagcca agcaaatgga
     2281 cgaatccact tggatcctga tcaagctgaa caaggcttag cgcagcttgt gatgacagtt
     2341 attgagctat tgaggcaaat cgttgaacgt catgccatga ggcgggtgga gggtggaacg
     2401 ttgacggacg aacaaattga aaacttagga attgcactaa tgaacttaga agaaaaaatg
     2461 gacgagttga aagaggtgtt cggtctggat gcagaagatt taaatattga tcttggaccg
     2521 ctaggcagcc tgctttaagc ggtcagtagg aggaacagta tggcagtcga acataatatg
     2581 cagtcaagta cgattgtaga tgtgctcgaa aagattttgg ataaaggagt cgttatagcg
     2641 ggggacatca ccgtaggaat tgcagatgtc gagctattaa cgataaagat ccgcttgatt
     2701 gtggcttcgg ttgataaggc aaaagaaatc ggcatggact ggtgggaaaa tgatccgtat
     2761 ctcagttcaa aaggagccaa taacaaagcg ctcgaagaag aaaataaaat gctgcatgag
     2821 cggttaaaaa cgcttgaaga aaaaatagaa acgaaacgtt aaaaactgta cgctacttaa
     2881 aaaatggagg gatttacaat ggcaactgaa acaaaattag ataacacaca ggcagaaaac
     2941 aaggaaaata aaaatgcgga aaacggttca aaagaaaaga acggttcaaa agcaagcaaa
     3001 acaacaagca gcgggccaat caaacgagcg gtagcaggag gcatcatcgg tgcaacgatt
     3061 ggatatgtat cgactcctga aaatcgaaaa agtctccttg accgcattga tacagacgaa
     3121 ttaaaaagca aagcatctga tttaggaaca aaggtaaaag aaaaatcaaa aagcagcgtg
     3181 gccagcctga aaacatctgc gggaagcttg tttaaaaaag ataaagataa atcaaaagat
     3241 gatgaagaaa acgtaaattc ttctagtagc gaaacagaag acgataacgt tcaagagtac
     3301 gacgagttaa aagaagaaaa tcaaactctt caagatcgct tatcacagct tgaagaaaaa
     3361 atgaacatgc ttgttgagct tagcctcaat aaaaatcaag acgaagaagc ggaagataca
     3421 gattccgacg aagaagagaa cgatgagaac gatgaaaacg atgaaaacga gcaggacgat
     3481 gaaaacgaag aagaaacatc taagccacgt aaaaaggata aaaaagaagc tgaggaagaa
     3541 gaaagtgaaa gtgacgaaga cagcgaggaa gaagaggaag attctcgctc aaacaaaaaa
     3601 aataaaaaag taaaaacaga agaagaagac gaagatgaaa gcgaagaaga aaaaaaggaa
     3661 gcgaaaccaa aaaagtcaac agctaaaaaa tcaaaaaata caaaagcaaa gaaaaacacg
     3721 gacgaagaag atgatgaagc aacatctctt tctagtgaag acgatacaac agcctaagac
     3781 gtaaaggagg aaagaaagac atgagtacag gcccttcttt ttcaactaaa gacaatacgc
     3841 ttgaatactt tgtgaaagct tctaataaac acggcttttc acttgatatt tcattaaatg
     3901 taaacggcgc tgtgatttcc ggtaccatga tttcagcaaa agaatacttt gattacttaa
     3961 gcgaaacgtt tgaagaaggc agtgaagtgg ctcaggcgct aagcgaacaa ttctctttag
     4021 caagcgaagc gagcgaatca aacggagaag cagaagccca ttttattcat ttgaaaaata
     4081 caaagattta ctgtggagac agtaaatcta ctccttctaa aggcaaaatc ttttggagag
     4141 ggaaaatagc agaagtagac gggtttttct taggaaagat ttctgatgca aaatcaacga
     4201 gtaaaaagag ttcataaaaa acggcggggt gattgccccg ccgtttttta gtgatgtgat
     4261 gagatgtgca gcttcttttt tcgatacatg agcggagtca tcttcatttc ttttcgaaac
     4321 tgattaataa aatagcttgt actattaaac cctacttgat aggcgacttc cgtcacattt
     4381 gcttctgctt gctgcagtcc ggattcgcga acgcgtctcg agagatccgg ctgctaacaa
     4441 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     4501 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccgacta
     4561 gtagcccgcc taatgagcgg gctttttttt aattccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4741 ttttgccttc ctgtttttgc tcacccagaa acgctcgtga aagtaaaaga cgcagaggac
     4801 caattggggg cacgagtggg atacatagaa ctggacttga atagcggtaa aatccttgag
     4861 agttttcgcc ctgaagagcg ttttccaatg atgagcactt tcaaagttct gctatgtgga
     4921 gcagtattat cccgtgtaga tgcggggcaa gagcaactcg gacgacgaat acactattcg
     4981 cagaatgact tggttgaata ctccccagtg acagaaaagc accttacgga cggaatgacg
     5041 gtaagagaat tatgtagtgc cgccataacg atgagtgata acactgcggc gaacttactt
     5101 ctgacaacca tcggtggacc gaaggaatta accgcttttt tgcacaatat gggagaccat
     5161 gtaactcgcc ttgaccgttg ggaaccagaa ctgaatgaag ccataccaaa cgacgagcga
     5221 gacaccacaa tgcctgcggc aatggcaaca acattacgca aactattaac tggcgaacta
     5281 cttactctgg cttcacggca acaattaata gactggcttg aagcggataa agttgcagga
     5341 ccactactgc gttcggcact tcctgctggc tggtttattg ctgataaatc tggggcagga
     5401 gagcgtggtt cacggggtat cattgccgca cttggaccag atggtaagcc ttcccgtatc
     5461 gtagttatct acacgacggg tagtcaggca actatggacg aacgaaatag acagattgct
     5521 gaaatagggg cttcactgat taagcattgg taaaccgata caattaaagg ctccttttgg
     5581 agcctttttt tttggacgga ccggtagaaa agatcaaagg atcttcttga gatccttttt
     5641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5761 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     6421 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     6481 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     6541 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     6601 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     6661 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     6721 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     6781 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     6841 tccgtgtaag ggggatttct gttcatgggg taatgatacc gatgaaacga gagaggatgc
     6901 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     6961 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     7021 tcgttaatac agatgtaggt gttggcacca tgcatcgatc ccgcgaaatt aatacgactc
     7081 actataggga gaccacaacg gtttccctct agagatagct tagcgatgtg aaagaagctg
     7141 gagaaaactt tcaagaaaaa gtttcttctt cagacgataa agagaagtca aaaaataagc
     7201 ggaaaataaa aggcgtaaat cagattaaaa aagcaacgga tattaaaagc tctactaaga
     7261 ttaaaagttc taatgatatt aaatcatcta cagatttaaa aacaatggga tcataagcga
     7321 aaggagaaat gagatatgtc tattcaaaaa agtactaata gttcaagttt agcagaagtc
     7381 attgaccgta ttttagataa aggaattgtt attgatgctt ttgcaagagt ttctgttgta
     7441 ggaattgaaa ttttaacgat tgaagcgcga gtggttattg ccagtgttga tacatggtta
     7501 cgctatgcag aagcagtagg gcttcttcgt gacgacgtag aagaaaacgg tcttcctgaa
     7561 cgttcaaatt caagtgaagg gcagccgcgt tttagtattt aatatgaagt ggtatggaac
     7621 cttccttttt tgagggtggt tccagctttc aggaggaacg taaaaatgac cgtcttaaca
     7681 gacaaaagga aaaaaggcag tggagctttt atacaagatg acgagacaaa agaggttctt
     7741 tcaagagcgc tgagctattt aaaatccggc tattccattc attttacagg tcctgccggc
     7801 ggaggcaaaa cctctttagc gcgagcgctt gctaaaaaga gaaagcgtcc tgtaatgctg
     7861 atgcacggga atcacgagct caacaacaaa gatttaattg gcgattttac gggatacacg
     7921 agcaaaaaag taatcgacca gtacgttcgt tctgtctata aaaaagatga acaggtgagt
     7981 gaaaactggc aggatggccg attgcttgaa gctgtaaaaa atggctatac gctgatttac
     8041 gacgaattta ctcgttctaa gcctgcgacg aataatatct ttctatcgat attagaagaa
     8101 ggcgtgctgc cgctgtatgg agtaaaaatg accgatcctt ttgtgcgcgt gcatcccgat
     8161 ttccgcgtca tcttcacaag caatccagct gagtatgccg gcgtatatga tacgcaagat
     8221 gcgcttctcg acaggttaat taccatgttt attgattata aagacatcga cagagagaca
     8281 gcgattttaa cggagaaaac ggacgtagaa gaagatgaag cgcgcacaat tgtaacgctc
     8341 gtagcaaacg tgcgaaaccg ctctggagac gaaaacagca gcggacttag cctgcgggct
     8401 tcgcttatga tcgctaccct tgccacgcag caagacattc ctatcgatgg aagtgac
//
