LOCUS       sequence-445017-        5555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Small BiT-Linker-GvpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5555)
  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 5555)
  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-R
FEATURES             Location/Qualifiers
     source          1..5555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     218..237
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        218..236
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    237..261
                     /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             276..298
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(738..756)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      752..799
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      836..1291
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(923..942)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1133..1154
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1383..2198)
                     /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(1383..2198)
                     /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     2106..2125
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2190..2209
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2320..2865
                     /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     2766..2785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3031..3048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3063..3202
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3128..3147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3288..3310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3304..3495)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4304..5386)
                     /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     5348..5367
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5387..5464)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
       61 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
      121 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
      181 tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa
      241 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
      301 tacatatggt gaccggctac cggctgttcg aggagattct ggtttctcaa ggcagttcag
      361 gtggtggcgg gagcggaggt ggaggctcga gcggtgaaat taaaaaaatt atgcaagccg
      421 tgaacgactt tttcggtgaa cacgtagctc ctcctcataa aattacctcg gtggaagcta
      481 ctgaagatga aggttggaga gttattgttg aagtcattga agaacgagaa tatatgaaaa
      541 aatacgccaa agatgaaatg ctcggaacgt acgagtgctt tgtaaataaa gaaaaagaag
      601 tcatttcatt caaacgactc gacgtcagat atagaagcgc cattggcatt gaagcataag
      661 gctgttttgg cggatgagag aagattgaga tccggctgct aacaaagccc gaaaggaagc
      721 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
      781 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
      841 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
      901 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
      961 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1021 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1081 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1141 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1201 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1261 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     1321 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     1381 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1441 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1501 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1561 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1621 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1681 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1741 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1801 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1861 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     1921 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1981 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2041 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     2101 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2161 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     2221 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     2281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgaaatcgtt ttggtctgcg
     2341 cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt cgaaggttct
     2401 ctgaactacc aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact
     2461 tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat
     2521 taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat
     2581 agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct
     2641 tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat
     2701 aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg
     2761 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag
     2821 cgtcagattt cgtgatgctt gtcagggggg cggagcctat ggaaaaacgt atggaaaaac
     2881 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2941 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3001 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3061 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     3121 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     3181 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     3241 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3301 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     3361 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     3421 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     3481 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     3541 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     3601 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3661 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3721 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3781 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3841 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3901 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     3961 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     4021 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     4081 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     4141 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     4201 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     4261 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     4321 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4381 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     4441 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     4501 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     4561 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     4621 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     4681 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     4741 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     4801 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     4861 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     4921 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     4981 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     5041 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     5101 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     5161 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     5221 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     5281 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     5341 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     5401 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     5461 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     5521 aggttgaggc cgttgagcac cgccgccgca aggaa
//
