LOCUS       sequence-445875-        6056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Small BiT-Linker-GvpF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  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 6056)
  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: pET26b(modified)-Sm-F
FEATURES             Location/Qualifiers
     source          1..6056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(407..1489)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1451..1470
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1490..1567)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1773..1792
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1876..1895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1876..1894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1895..1919
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1934..1956
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2614..2631
                     /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(2897..2915)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2911..2958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2995..3450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3082..3101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3292..3313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3542..4357)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3542..4357)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4265..4284
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4349..4368
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4479..5024
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4925..4944
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5190..5207
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5222..5361
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5287..5306)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5447..5469
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5463..5654)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
       61 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
      121 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
      181 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
      241 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
      301 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
      361 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
      421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      481 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
      541 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
      601 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
      661 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
      721 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
      781 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
      841 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
      901 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
      961 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     1021 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     1081 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     1141 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     1201 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     1261 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     1321 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     1381 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     1441 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     1501 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     1561 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     1621 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     1681 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     1741 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     1801 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     1861 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     1921 ctctagaaat aattttgttt aactttaaga aggagatata catatggtga ccggctaccg
     1981 gctgttcgag gagattctgg tttctcaagg cagttcaggt ggtggcggga gcggaggtgg
     2041 aggctcgagc ggtagtgaaa caaacgaaac aggtatttat atttttagcg ccattcaaac
     2101 ggataaagac gaagaatttg gcgccgtgga agtagaagga acaaaagctg aaacattttt
     2161 gattcgctac aaagacgcgg ctatggtagc agctgaagta ccgatgaaaa tttatcatcc
     2221 taatcgccaa aatttattaa tgcatcaaaa cgcagtagca gcgattatgg acaagaacga
     2281 tacggttatt ccaatcagct ttgggaatgt attcaaatca aaagaagacg taaaagttct
     2341 tttggaaaac ctttatccgc agtttgaaaa gctgtttcca gcgatcaaag gaaaaattga
     2401 agtcggttta aaagtaattg ggaaaaaaga atggcttgag aaaaaagtaa acgaaaatcc
     2461 tgaacttgag aaagtatcag catccgtaaa aggaaaatca gaagcagccg gttattatga
     2521 gcgtattcaa cttggaggaa tggctcaaaa gatgtttact tccctgcaaa aagaagtcaa
     2581 gacagatgtg ttttctccgc ttgaagaagc agcggaagca gcaaaagcaa atgagccaac
     2641 gggcgaaacg atgcttttaa acgcgtcttt cttaattaac cgagaagatg aagcgaagtt
     2701 tgatgaaaaa gtaaatgaag cgcatgaaaa ctggaaagac aaagccgatt ttcattacag
     2761 cggtccttgg cctgcttata attttgtgaa cattcgccta aaagtagaag agaaataagg
     2821 ctgttttggc ggatgagaga agattgagat ccggctgcta acaaagcccg aaaggaagct
     2881 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     2941 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     3001 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3061 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3121 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3181 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3241 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3301 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3361 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3421 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     3481 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt
     3541 cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     3601 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     3661 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     3721 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     3781 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     3841 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     3901 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     3961 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     4021 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     4081 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     4141 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     4201 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     4261 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     4321 gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     4381 tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4441 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatcgttt tggtctgcgc
     4501 gtaatctctt gctctgaaaa cggaaaaacc gccttgcagg gcggtttttc gaaggttctc
     4561 tgaactacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt
     4621 gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt
     4681 accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata
     4741 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt
     4801 ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata
     4861 acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc
     4921 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
     4981 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacgta tggaaaaacg
     5041 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5101 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5161 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5221 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     5281 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     5341 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5401 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5461 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     5521 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     5581 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     5641 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     5701 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     5761 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     5821 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     5881 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     5941 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     6001 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaacc
//
