LOCUS       sequence-445019-        6054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GvpF-Linker-Small BiT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6054)
  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 6054)
  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)-F-Sm
FEATURES             Location/Qualifiers
     source          1..6054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             412..603
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(597..619)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    705..844
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     760..779
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(859..876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1042..1586)
                     /direction=LEFT
                     /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     complement(1122..1141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1697..1716)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1708..2523
                     /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             1708..2523
                     /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     complement(1781..1800)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2615..3070)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2752..2773)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2964..2983
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3107..3154)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3150..3168
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3521..3538)
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="GSSKSK"
     RBS             4109..4131
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4146..4170
                     /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)."
     primer_bind     complement(4170..4189)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4171..4189)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4273..4292)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4498..4575
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4576..5658
                     /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     complement(4595..4614)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc
       61 tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa
      121 agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga
      181 tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc
      241 tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt
      301 tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc
      361 atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa
      421 caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg
      481 gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac
      541 cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
      601 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
      661 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
      721 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
      781 tgagagtgca ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      841 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      901 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      961 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1021 gctggcgttt ttccatacgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     1081 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1141 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     1201 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     1261 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     1321 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     1381 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     1441 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     1501 gttcagagaa ccttcgaaaa accgccctgc aaggcggttt ttcgttttca gagcaagaga
     1561 ttacgcgcag accaaaacga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct
     1681 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct
     1741 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1801 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     1861 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     1921 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     1981 gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa
     2041 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2101 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2161 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2221 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2281 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2341 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2401 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2461 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2521 taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2581 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     2641 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2701 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2761 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2821 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2881 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2941 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3001 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3061 cagggcgcgt cccattcgcc aatccggata tagttcctcc tttcagcaaa aaacccctca
     3121 agacccgttt agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca
     3181 actcagcttc ctttcgggct ttgttagcag ccggatctca atcttctctc atccgccaaa
     3241 acagcctcac agaatctcct cgaacagccg gtagccggtc acaccgctcg agcctccacc
     3301 tccgctcccg ccaccacctg aactgccttg agaaactttc tcttctactt ttaggcgaat
     3361 gttcacaaaa ttataagcag gccaaggacc gctgtaatga aaatcggctt tgtctttcca
     3421 gttttcatgc gcttcattta ctttttcatc aaacttcgct tcatcttctc ggttaattaa
     3481 gaaagacgcg tttaaaagca tcgtttcgcc cgttggctca tttgcttttg ctgcttccgc
     3541 tgcttcttca agcggagaaa acacatctgt cttgacttct ttttgcaggg aagtaaacat
     3601 cttttgagcc attcctccaa gttgaatacg ctcataataa ccggctgctt ctgattttcc
     3661 ttttacggat gctgatactt tctcaagttc aggattttcg tttacttttt tctcaagcca
     3721 ttcttttttc ccaattactt ttaaaccgac ttcaattttt cctttgatcg ctggaaacag
     3781 cttttcaaac tgcggataaa ggttttccaa aagaactttt acgtcttctt ttgatttgaa
     3841 tacattccca aagctgattg gaataaccgt atcgttcttg tccataatcg ctgctactgc
     3901 gttttgatgc attaataaat tttggcgatt aggatgataa attttcatcg gtacttcagc
     3961 tgctaccata gccgcgtctt tgtagcgaat caaaaatgtt tcagcttttg ttccttctac
     4021 ttccacggcg ccaaattctt cgtctttatc cgtttgaatg gcgctaaaaa tataaatacc
     4081 tgtttcgttt gtttcactca tatgtatatc tccttcttaa agttaaacaa aattatttct
     4141 agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga
     4201 tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     4261 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     4321 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     4381 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
     4441 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac
     4501 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     4561 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     4621 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     4681 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     4741 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     4801 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     4861 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     4921 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     4981 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     5041 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     5101 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     5161 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     5221 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     5281 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     5341 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     5401 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     5461 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     5521 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     5581 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5641 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc
     5701 gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     5761 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     5821 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat
     5881 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac
     5941 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggcccc
     6001 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc gggg
//
