LOCUS       sequence-180859-        8764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Mega GVs in E.Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8764)
  AUTHORS   Lakshmanan A, Lu GJ, Farhadi A, Nety SP, Kunth M, Lee-Gosselin A, 
            Maresca D, Bourdeau RW, Yin M, Yan J, Witte C, Malounda D, Foster 
            FS, Schroder L, Shapiro MG
  TITLE     Preparation of biogenic gas vesicle nanostructures for use as 
            contrast agents for ultrasound and MRI.
  JOURNAL   Nat Protoc. 2017 Oct;12(10):2050-2080. doi: 10.1038/nprot.2017.081. 
            Epub 2017 Sep 7.
  PUBMED    28880278
REFERENCE   2  (bases 1 to 8764)
  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/nprot.2017.081"; journalName: "Nat Protoc"; date: 
            "2017-10"; volume: "12"; issue: "10"; pages: "2050-2080"
COMMENT     Sequence Label: pST39-pNL29
FEATURES             Location/Qualifiers
     source          1..8764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2408..2425
                     /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(6168..6186)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6182..6229
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6296..6387
                     /gene="bla"
                     /label=AmpR promoter
     terminator      7252..7291
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      7322..7910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7811..7830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8064..8081
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8096..8235
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8161..8180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8321..8343
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8337..8528)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagagatagc ttagcgatgt
       61 gaaagaagct ggagaaaact ttcaagaaaa agtttcttct tcagacgata aagagaagtc
      121 aaaaaataag cggaaaataa aaggcgtaaa tcagattaaa aaagcaacgg atattaaaag
      181 ctctactaag attaaaagtt ctaatgatat taaatcatct acagatttaa aaacaatggg
      241 atcataagcg aaaggagaaa tgagatatgt ctattcaaaa aagtactaat agttcaagtt
      301 tagcagaagt cattgaccgt attttagata aaggaattgt tattgatgct tttgcaagag
      361 tttctgttgt aggaattgaa attttaacga ttgaagcgcg agtggttatt gccagtgttg
      421 atacatggtt acgctatgca gaagcagtag ggcttcttcg tgacgacgta gaagaaaacg
      481 gtcttcctga acgttcaaat tcaagtgaag ggcagccgcg ttttagtatt taatatgaag
      541 tggtatggaa ccttcctttt ttgagggtgg ttccagcttt tagatctaac tattggaggc
      601 tactaaaaat ggaaattaaa aaaattatgc aagccgtgaa cgactttttc ggtgaacacg
      661 tagctcctcc tcataaaatt acctcggtgg aagctactga agatgaaggt tggagagtta
      721 ttgttgaagt cattgaagaa cgagaatata tgaaaaaata cgccaaagat gaaatgctcg
      781 gaacgtacga gtgctttgta aataaagaaa aagaagtcat ttcattcaaa cgactcgacg
      841 tcagatatag aagcgccatt ggcattgaag cataaacgat aagatggcag gaggaacgta
      901 aaaatgaccg tcttaacaga caaaaggaaa aaaggcagtg gagcttttat acaagatgac
      961 gagacaaaag aggttctttc aagagcgctg agctatttaa aatccggcta ttccattcat
     1021 tttacaggtc ctgccggcgg aggcaaaacc tctttagcgc gagcgcttgc taaaaagaga
     1081 aagcgtcctg taatgctgat gcacgggaat cacgagctca acaacaaaga tttaattggc
     1141 gattttacgg gatacacgag caaaaaagta atcgaccagt acgttcgttc tgtctataaa
     1201 aaagatgaac aggtgagtga aaactggcag gatggccgat tgcttgaagc tgtaaaaaat
     1261 ggctatacgc tgatttacga cgaatttact cgttctaagc ctgcgacgaa taatatcttt
     1321 ctatcgatat tagaagaagg cgtgctgccg ctgtatggag taaaaatgac cgatcctttt
     1381 gtgcgcgtgc atcccgattt ccgcgtcatc ttcacaagca atccagctga gtatgccggc
     1441 gtatatgata cgcaagatgc gcttctcgac aggttaatta ccatgtttat tgattataaa
     1501 gacatcgaca gagagacagc gattttaacg gagaaaacgg acgtagaaga agatgaagcg
     1561 cgcacaattg taacgctcgt agcaaacgtg cgaaaccgct ctggagacga aaacagcagc
     1621 ggacttagcc tgcgggcttc gcttatgatc gctacccttg ccacgcagca agacattcct
     1681 atcgatggaa gtgacgaaga ttttcaaacg ttatgtatcg atattttgca tcatccgctt
     1741 accaaatgtt tggatgaaga aaatgcaaaa agcaaagccg aaaaaatcat tttagaagaa
     1801 tgtaagaata tagacactga agaaaagtaa aggagcttga aaacatgagt gaaacaaacg
     1861 aaacaggtat ttatattttt agcgccattc aaacggataa agacgaagaa tttggcgccg
     1921 tggaagtaga aggaacaaaa gctgaaacat ttttgattcg ctacaaagac gcggctatgg
     1981 tagcagctga agtaccgatg aaaatttatc atcctaatcg ccaaaattta ttaatgcatc
     2041 aaaacgcagt agcagcgatt atggacaaga acgatacggt tattccaatc agctttggga
     2101 atgtattcaa atcaaaagaa gacgtaaaag ttcttttgga aaacctttat ccgcagtttg
     2161 aaaagctgtt tccagcgatc aaaggaaaaa ttgaagtcgg tttaaaagta attgggaaaa
     2221 aagaatggct tgagaaaaaa gtaaacgaaa atcctgaact tgagaaagta tcagcatccg
     2281 taaaaggaaa atcagaagca gccggttatt atgagcgtat tcaacttgga ggaatggctc
     2341 aaaagatgtt tacttccctg caaaaagaag tcaagacaga tgtgttttct ccgcttgaag
     2401 aagcagcgga agcagcaaaa gcaaatgagc caacgggcga aacgatgctt ttaaacgcgt
     2461 ctttcttaat taaccgagaa gatgaagcga agtttgatga aaaagtaaat gaagcgcatg
     2521 aaaactggaa agacaaagcc gattttcatt acagcggtcc ttggcctgct tataattttg
     2581 tgaacattcg cctaaaagta gaagagaaat aacgtgcttc acaaattagt aaccgcaccc
     2641 attaaccttg tagtgaaaat cggcgaaaaa gtacaggaag aagctgataa acagctatat
     2701 gaccttccga cgattcagca aaagctcatt cagcttcaaa tgatgtttga gcttggtgaa
     2761 attccagaag aagcgtttca agaaaaagaa gatgaattgt taatgaggta cgaaattgcg
     2821 aaacgcagag aaattgaaca atgggaagag ctaacacaaa aaagaaatga ggaatcctag
     2881 atgggagaat tactgtattt atacggttta attccaacaa aagaagcagc agccatagag
     2941 ccgtttccat cttataaggg gtttgacgga gaacattcac tgtacccaat tgcgtttgat
     3001 caggtgacgg ctgtagtttc taagctggat gctgacacct attcagaaaa agtgattcaa
     3061 gaaaaaatgg agcaggatat gagctggctg caggaaaaag catttcatca tcacgaaacg
     3121 gtagccgctt tgtacgaaga atttacgatc attccattaa aattttgcac catttataaa
     3181 ggtgaagaaa gtctgcaggc agctattgag attaacaaag aaaagataga gaattcactg
     3241 acgctgcttc aaggaaatga agagtggaat gtgaaaattt actgtgatga tacagagctt
     3301 aaaaaaggaa tcagcgaaac gaatgaaagc gtgaaagcga aaaaacaaga aattagtcac
     3361 ttatcaccag gaagacagtt ttttgaaaag aaaaaaatag atcagctgat tgaaaaagaa
     3421 ttagagcttc acaaaaacaa agtgtgtgaa gagatacatg acaagctaaa agaattatcg
     3481 ctttatgact ctgttaaaaa gaattggagc aaagacgtaa ctggcgcagc tgaacagatg
     3541 gcgtggaaca gcgtgtttct tctcccgtct ctgcagatta ctaagttcgt aaacgaaata
     3601 gaagagcttc agcaaaggct tgaaaataaa ggctggaagt ttgaagtgac gggaccatgg
     3661 ccgccctatc atttctcgag ctttgcgtaa agtgaggaat taacattatg tctcttaaac
     3721 aatccatgga gaataaagat attgctctta ttgatatttt agatgtcatt ttagataaag
     3781 gagtcgccat taaaggagac ttaatcattt ccatagctgg cgtcgattta gtgtatttgg
     3841 atttgcgggt gcttatttct tcggttgaaa cgcttgtgca agcaaaagaa ggaaatcaca
     3901 aaccaatcac ttctgaacaa tttgataaac aaaaggagga attaatggat gcaaccggtc
     3961 agccaagcaa atggacgaat ccacttggat cctgatcaag ctgaacaagg cttagcgcag
     4021 cttgtgatga cagttattga gctattgagg caaatcgttg aacgtcatgc catgaggcgg
     4081 gtggagggtg gaacgttgac ggacgaacaa attgaaaact taggaattgc actaatgaac
     4141 ttagaagaaa aaatggacga gttgaaagag gtgttcggtc tggatgcaga agatttaaat
     4201 attgatcttg gaccgctagg cagcctgctt taagcggtca gtaggaggaa cagtatggca
     4261 gtcgaacata atatgcagtc aagtacgatt gtagatgtgc tcgaaaagat tttggataaa
     4321 ggagtcgtta tagcggggga catcaccgta ggaattgcag atgtcgagct attaacgata
     4381 aagatccgct tgattgtggc ttcggttgat aaggcaaaag aaatcggcat ggactggtgg
     4441 gaaaatgatc cgtatctcag ttcaaaagga gccaataaca aagcgctcga agaagaaaat
     4501 aaaatgctgc atgagcggtt aaaaacgctt gaagaaaaaa tagaaacgaa acgttaaaaa
     4561 ctgtacgcta cttaaaaaat ggagggattt acaatggcaa ctgaaacaaa attagataac
     4621 acacaggcag aaaacaagga aaataaaaat gcggaaaacg gttcaaaaga aaagaacggt
     4681 tcaaaagcaa gcaaaacaac aagcagcggg ccaatcaaac gagcggtagc aggaggcatc
     4741 atcggtgcaa cgattggata tgtatcgact cctgaaaatc gaaaaagtct ccttgaccgc
     4801 attgatacag acgaattaaa aagcaaagca tctgatttag gaacaaaggt aaaagaaaaa
     4861 tcaaaaagca gcgtggccag cctgaaaaca tctgcgggaa gcttgtttaa aaaagataaa
     4921 gataaatcaa aagatgatga agaaaacgta aattcttcta gtagcgaaac agaagacgat
     4981 aacgttcaag agtacgacga gttaaaagaa gaaaatcaaa ctcttcaaga tcgcttatca
     5041 cagcttgaag aaaaaatgaa catgcttgtt gagcttagcc tcaataaaaa tcaagacgaa
     5101 gaagcggaag atacagattc cgacgaagaa gagaacgatg agaacgatga aaacgatgaa
     5161 aacgagcagg acgatgaaaa cgaagaagaa acatctaagc cacgtaaaaa ggataaaaaa
     5221 gaagctgagg aagaagaaag tgaaagtgac gaagacagcg aggaagaaga ggaagattct
     5281 cgctcaaaca aaaaaaataa aaaagtaaaa acagaagaag aagacgaaga tgaaagcgaa
     5341 gaagaaaaaa aggaagcgaa accaaaaaag tcaacagcta aaaaatcaaa aaatacaaaa
     5401 gcaaagaaaa acacggacga agaagatgat gaagcaacat ctctttctag tgaagacgat
     5461 acaacagcct aagacgtaaa ggaggaaaga aagacatgag tacaggccct tctttttcaa
     5521 ctaaagacaa tacgcttgaa tactttgtga aagcttctaa taaacacggc ttttcacttg
     5581 atatttcatt aaatgtaaac ggcgctgtga tttccggtac catgatttca gcaaaagaat
     5641 actttgatta cttaagcgaa acgtttgaag aaggcagtga agtggctcag gcgctaagcg
     5701 aacaattctc tttagcaagc gaagcgagcg aatcaaacgg agaagcagaa gcccatttta
     5761 ttcatttgaa aaatacaaag atttactgtg gagacagtaa atctactcct tctaaaggca
     5821 aaatcttttg gagagggaaa atagcagaag tagacgggtt tttcttagga aagatttctg
     5881 atgcaaaatc aacgagtaaa aagagttcat aaaaaacggc ggggtgattg ccccgccgtt
     5941 ttttagtgat gtgatgagat gtgcagcttc ttttttcgat acatgagcgg agtcatcttc
     6001 atttcttttc gaaactgatt aataaaatag cttgtactat taaaccctac ttgataggcg
     6061 acttccgtca catttgcttc tgcttgctgc agtccggatt cgcgaacgcg tctcgagaga
     6121 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     6181 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     6241 aactatatcc gactagtagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt
     6301 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6361 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6421 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta
     6481 aaagacgcag aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc
     6541 ggtaaaatcc ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa
     6601 gttctgctat gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga
     6661 cgaatacact attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt
     6721 acggacggaa tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact
     6781 gcggcgaact tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac
     6841 aatatgggag accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata
     6901 ccaaacgacg agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta
     6961 ttaactggcg aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg
     7021 gataaagttg caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat
     7081 aaatctgggg caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt
     7141 aagccttccc gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga
     7201 aatagacaga ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt
     7261 aaaggctcct tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt
     7321 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7381 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7441 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     7501 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7561 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7621 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7681 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7741 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7801 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7861 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7921 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7981 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     8041 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     8101 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     8161 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     8221 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     8281 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     8341 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     8401 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     8461 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     8521 ttggtcactg atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga
     8581 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     8641 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     8701 gtcaatgcca gcgcttcgtt aatacagatg taggtgttgg caccatgcat cgatcccgcg
     8761 aaat
//
