LOCUS       sequence-346300-        6267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SynZip1 linked surfactant protein that localizes to RGG condensate 
            interface.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  AUTHORS   Guan M, Garabedian MV, Leutenegger M, Schuster BS, Good MC, Hammer 
            DA
  TITLE     Incorporation and Assembly of a Light-Emitting Enzymatic Reaction 
            into Model Protein Condensates.
  JOURNAL   Biochemistry. 2021 Oct 14. doi: 10.1021/acs.biochem.1c00373.
  PUBMED    34648259
REFERENCE   2  (bases 1 to 6267)
  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"; journalName: 
            "Biochemistry. 2021 Oct 14. doi: 10.1021/acs.biochem.1c00373."
COMMENT     Sequence Label: SZ1-GST-mCherry-RGG
FEATURES             Location/Qualifiers
     source          1..6267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             202..219
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(272..290)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      286..333
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      370..825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(457..476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     667..688
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(940..1755)
                     /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(940..1755)
                     /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     1663..1682
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1747..1766
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1908..2496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2397..2416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2650..2667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(2838..3920)
                     /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     3882..3901
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3921..3998)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4204..4223
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4307..4326
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4307..4325
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4326..4350
                     /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             4365..4387
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4543..5193
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
     primer_bind     5154..5176
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             5923..5943
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ggcggtggcg gtggcaatcg tggctataac aataaccgtg gtggcggtgg cggtggctac
       61 aatcgccagg atcgtggtga cggtggcagc tctaactttt cgcgcggtgg ctataataac
      121 cgtgatgaag gcagcgacaa ccgcggtagt ggccgttcct acaataacga tcgtcgcgac
      181 aatggtggcg atggcctcga gcaccaccac caccaccact gagatccggc tgctaacaaa
      241 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
      301 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
      361 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
      421 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
      481 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
      541 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
      601 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
      661 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
      721 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
      781 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
      841 gggtacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      901 taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat
      961 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     1021 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     1081 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     1141 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     1201 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     1261 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     1321 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     1381 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     1441 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     1501 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     1561 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     1621 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     1681 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg
     1741 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     1801 tcatgaccaa aatcccttaa cgtgacctag gtcatgacca aaatccctta acgtgagttt
     1861 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1921 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1981 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2041 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2101 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2161 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2221 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2281 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2341 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2401 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2461 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2521 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2581 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2641 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gccataatcc atgataaaga
     2701 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     2761 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     2821 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2881 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     2941 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     3001 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     3061 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     3121 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     3181 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     3241 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     3301 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     3361 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     3421 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     3481 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     3541 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     3601 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     3661 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     3721 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     3781 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     3841 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     3901 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     3961 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     4021 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     4081 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     4141 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     4201 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     4261 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     4321 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
     4381 aaggagatat acataatgaa cttggtagcc cagcttgaga acgaagttgc gagtctggaa
     4441 aacgaaaacg agacgttgaa gaaaaagaac ttgcacaaga aagatttgat tgcctacttg
     4501 gaaaaggaaa ttgccaatct gcgtaaaaag attgaggaat cgatgtcccc tatactaggt
     4561 tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa
     4621 aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt
     4681 gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca
     4741 cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca
     4801 aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt
     4861 tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta
     4921 cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat
     4981 catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac
     5041 ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc
     5101 ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg
     5161 caagccacgt ttggtggtgg cgaccatcct ccaggatccg gtatggtgag caagggcgag
     5221 gaggacaaca tggcgatcat caaggagttt atgcggttca aggtgcacat ggaaggttct
     5281 gtcaatggac acgaatttga aatcgagggt gagggggaag gacgcccata cgaggggacc
     5341 caaactgcga agcttaaagt tacaaagggg ggtccgttgc cgtttgcttg ggatatctta
     5401 tcaccgcagt tcatgtatgg cagcaaagct tacgtcaaac acccagcaga tattcctgat
     5461 tatcttaaat tgtcgttccc cgaaggcttc aagtgggaac gggtaatgaa cttcgaagat
     5521 ggaggggtag ttacggtcac ccaggattct tctctgcaag atggcgagtt catttacaaa
     5581 gtgaaactta gaggcacaaa tttcccctct gatggacccg ttatgcagaa gaaaacgatg
     5641 gggtgggagg catctagtga gcgtatgtat ccagaagatg gggctttaaa aggggagatc
     5701 aaacagcgtc tgaagttgaa agacggcggt cattatgatg ccgaggtcaa aactacgtac
     5761 aaggcaaaaa agccagttca gttgcctggt gcttacaacg tgaatatcaa acttgatatc
     5821 acaagccaca atgaggatta taccattgtt gaacaatatg aaagagcaga gggccggcac
     5881 agtacaggcg gtatggatga gttgtataaa ggcggcggat ctgagaattt atatttccag
     5941 ggcgaattcg ggaagcttat ggaatccaat caatccaata acggtggctc gggtaatgcc
     6001 gctctgaatc gtggtggccg ctacgtgccg ccgcatctgc gtggtggcga tggtggtgca
     6061 gcagcagctg catctgctgg cggtgatgac cgtcgcggcg gtgcgggcgg tggcggttat
     6121 cgtcgcggcg gtggcaatag tggtggcggt ggcggtggcg gttatgatcg tggttacaat
     6181 gacaaccgtg atgaccgcga taaccgtggc ggttccggcg gttacggtcg tgatcgtaat
     6241 tatgaagacc gtggttacaa cggcggt
//
