LOCUS       sequence-445014-        5573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Small BiT-Linker-GvpK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5573)
  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 5573)
  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-K
FEATURES             Location/Qualifiers
     source          1..5573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(649..1731)
                     /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     1693..1712
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1732..1809)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2015..2034
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2118..2137
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2118..2136
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2137..2161
                     /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             2176..2198
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2656..2674)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2670..2717
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2754..3209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2841..2860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3051..3072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3301..4116)
                     /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(3301..4116)
                     /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     4024..4043
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4108..4127
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4238..4783
                     /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     4684..4703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4949..4966
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4981..5120
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5046..5065)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5206..5228
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5222..5413)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
       61 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
      121 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
      181 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
      241 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
      301 acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
      361 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
      421 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
      481 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
      541 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
      601 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
      661 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      721 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      781 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      841 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      901 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
      961 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     1021 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     1081 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     1141 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     1201 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     1261 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     1321 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     1381 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     1441 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     1501 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     1561 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1621 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1681 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1741 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1801 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1861 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1921 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1981 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     2041 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     2101 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     2161 ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatggt gaccggctac
     2221 cggctgttcg aggagattct ggtttctcaa ggcagttcag gtggtggcgg gagcggaggt
     2281 ggaggctcga gcggtcaacc ggtcagccaa gcaaatggac gaatccactt ggatcctgat
     2341 caagctgaac aaggcttagc gcagcttgtg atgacagtta ttgagctatt gaggcaaatc
     2401 gttgaacgtc atgccatgag gcgggtggag ggtggaacgt tgacggacga acaaattgaa
     2461 aacttaggaa ttgcactaat gaacttagaa gaaaaaatgg acgagttgaa agaggtgttc
     2521 ggtctggatg cagaagattt aaatattgat cttggaccgc taggcagcct gctttaaggc
     2581 tgttttggcg gatgagagaa gattgagatc cggctgctaa caaagcccga aaggaagctg
     2641 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     2701 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
     2761 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2821 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2881 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2941 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3001 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3061 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3121 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3181 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     3241 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     3301 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     3361 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     3421 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     3481 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     3541 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     3601 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     3661 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     3721 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     3781 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     3841 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     3901 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     3961 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     4021 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     4081 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     4141 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     4201 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcgtttt ggtctgcgcg
     4261 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
     4321 gaactaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
     4381 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
     4441 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
     4501 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
     4561 gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa
     4621 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
     4681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     4741 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacgtat ggaaaaacgc
     4801 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4861 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4921 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4981 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     5041 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     5101 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5161 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5221 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     5281 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     5341 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     5401 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     5461 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     5521 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag aga
//
