LOCUS       sequence-445898-        6167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GvpT-Linker-Small BiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6167)
  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 6167)
  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)-T-Sm
FEATURES             Location/Qualifiers
     source          1..6167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(100..119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     310..331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(560..1375)
                     /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(560..1375)
                     /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     1283..1302
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1367..1386
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1497..2042
                     /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     1943..1962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2208..2225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2240..2379
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2305..2324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2465..2487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2481..2672)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3481..4563)
                     /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     4525..4544
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4564..4641)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4847..4866
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4950..4969
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4950..4968
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4969..4993
                     /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             5008..5030
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6082..6100)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6096..6143
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
       61 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      121 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      181 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
      241 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      301 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
      361 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
      421 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc attgcagggc
     1561 ggtttttcga aggttctctg aactaccaac tctttgaacc gaggtaactg gcttggagga
     1621 gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact
     1681 aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg
     1741 gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc
     1801 gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat
     1861 gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag
     1921 agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1981 gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga
     2041 aaaacgtatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2101 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2161 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2221 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2281 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2341 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2401 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2461 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2521 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     2581 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     2641 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     2701 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     2761 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     2821 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     2881 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     2941 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3001 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3061 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     3121 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     3181 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     3241 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     3301 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     3361 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     3421 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     3481 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3541 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     3601 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     3661 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     3721 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     3781 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     3841 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     3901 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     3961 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     4021 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     4081 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     4141 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     4201 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     4261 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     4321 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     4381 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     4441 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     4501 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     4561 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     4621 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     4681 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     4741 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     4801 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     4861 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     4921 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     4981 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     5041 gcaactgaaa caaaattaga taacacacag gcagaaaaca aggaaaataa aaatgcggaa
     5101 aacggttcaa aagaaaagaa cggttcaaaa gcaagcaaaa caacaagcag cgggccaatc
     5161 aaacgagcgg tagcaggagg catcatcggt gcaacgattg gatatgtatc gactcctgaa
     5221 aatcgaaaaa gtctccttga ccgcattgat acagacgaat taaaaagcaa agcatctgat
     5281 ttaggaacaa aggtaaaaga aaaatcaaaa agcagcgtgg ccagcctgaa aacatctgcg
     5341 ggaagcttgt ttaaaaaaga taaagataaa tcaaaagatg atgaagaaaa cgtaaattct
     5401 tctagtagcg aaacagaaga cgataacgtt caagagtacg acgagttaaa agaagaaaat
     5461 caaactcttc aagatcgctt atcacagctt gaagaaaaaa tgaacatgct tgttgagctt
     5521 agcctcaata aaaatcaaga cgaagaagcg gaagatacag attccgacga agaagagaac
     5581 gatgagaacg atgaaaacga tgaaaacgag caggacgatg aaaacgaaga agaaacatct
     5641 aagccacgta aaaaggataa aaaagaagct gaggaagaag aaagtgaaag tgacgaagac
     5701 agcgaggaag aagaggaaga ttctcgctca aacaaaaaaa ataaaaaagt aaaaacagaa
     5761 gaagaagacg aagatgaaag cgaagaagaa aaaaaggaag cgaaaccaaa aaagtcaaca
     5821 gctaaaaaat caaaaaatac aaaagcaaag aaaaacacgg acgaagaaga tgatgaagca
     5881 acatctcttt ctagtgaaga cgatacaaca gccgtttctc aaggcagttc aggtggtggc
     5941 gggagcggag gtggaggctc gagcggtgtg accggctacc ggctgttcga ggagattctg
     6001 tgaggctgtt ttggcggatg agagaagatt gagatccggc tgctaacaaa gcccgaaagg
     6061 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     6121 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccgga
//
