LOCUS       sequence-437136-        7769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tobacco etch virus protease-responsive elastin-like polypeptide 
            (ELP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7769)
  AUTHORS   Wirtz BM, Yun AG, Wick C, Gao XJ, Mai DJ
  TITLE     Protease-Driven Phase Separation of Elastin-Like Polypeptides.
  JOURNAL   Biomacromolecules. 2024 Jul 9. doi: 10.1021/acs.biomac.4c00346.
  PUBMED    38980747
REFERENCE   2  (bases 1 to 7769)
  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: 
            "Biomacromolecules. 2024 Jul 9. doi: 10.1021/acs.biomac.4c00346."
COMMENT     Sequence Label: pET22b-TEVP-cleavableELP
FEATURES             Location/Qualifiers
     source          1..7769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..24
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             946..963
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1016..1034)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1030..1077
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1114..1569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1201..1220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1411..1432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1595..1699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1700..1768,1769..2560)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1918..1937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2731..3319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3220..3239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3473..3490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3505..3644
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3570..3589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3730..3752
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3746..3937)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4746..5828)
                     /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     5790..5809
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5829..5906)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6112..6131
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6215..6234
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6215..6233
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6234..6258
                     /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             6273..6295
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ggtgagaact tatactttca gagcggtggc ggctccgtgc caggcgtcgg tgtgcctgga
       61 gtcggggtgc caggggtagg agtccctggc gtaggagtgc caggagtggg tgttccgggt
      121 gtaggagttc ctggagtggg ggttcccggt gttggcgtac cgggcgttgg agtacctggt
      181 gtaggtgtgc ccggtgtagg cgtgcctggc gttggtgttc caggagttgg ggtccccgga
      241 gttggcgttc caggtgtcgg ggttccgggg gttggagtgc ctggtgtggg cgtacctgga
      301 gtaggagtac ccggtgtggg agtgccggga gttggagtcc caggggtcgg cgttcctggg
      361 gtgggagtcc cgggggtggg cgttcccgga gttggggtgc ccggagtagg cgtaccaggc
      421 gtgggagtac cgggtgtggg ggtcccaggt gtaggggtac ccggcgtggg tgtgccgggt
      481 gtcggagttc ccggagtcgg agtccccggg gtgggtgtac cgggggtcgg ggtaccaggg
      541 gtgggggtgc cgggggtagg cgttccgggc gtaggggttc ctggtgtcgg tgttcccggc
      601 gtaggtgttc ctggcgtggg ggtaccggga gtgggcgtcc cgggtgttgg agttccggga
      661 gtaggtgtac ccggagtggg agttccaggc gttggggttc caggggttgg cgtcccagga
      721 gtaggtgtac ctggggtagg tgtccctggt gttggtgtac ctggcgtcgg agtgcccggg
      781 gttggggtgc ctggggttgg tgtgccaggt gttggggtac ctggggtcgg agtaccagga
      841 gtcggtgtac caggtgtggg tgtcccaggc gtaggcgtcc ctggagttgg tgtccccggt
      901 gtcggcgtac ccggggtcgg tgtcccggga gtcggctggc tcgagcacca ccaccaccac
      961 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1021 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     1081 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     1141 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1201 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1261 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1321 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1381 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     1441 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1501 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     1561 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1621 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1681 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1741 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1801 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1861 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1921 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1981 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2041 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2101 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2161 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2221 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2281 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2341 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2401 agccggtgag cgtggctctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2461 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2521 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2581 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2641 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2701 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2761 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2821 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2881 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2941 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3001 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3061 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3481 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3541 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3601 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3661 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3721 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3781 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3841 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3901 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3961 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     4021 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     4081 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     4141 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     4201 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     4261 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     4321 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4381 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     4441 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     4501 ataccgcaag cgacaggccg atcatcgtcg cgcgccagcg aaagcggtcc tcgccgaaaa
     4561 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     4621 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     4681 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4741 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4801 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4861 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     4921 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     4981 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     5041 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     5101 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     5161 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     5221 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     5281 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     5341 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5401 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5461 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     5521 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     5581 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     5641 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     5701 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5761 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5821 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     5881 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     5941 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     6001 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     6061 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6121 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6181 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg
     6241 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     6301 atatgggtgg ttggggacca gtgcctggcg tcggtgtacc tggtgctggt gtacccggtg
     6361 ctggagtacc gggggcaggt gtaccaggtg caggggtccc aggagctgga gttccaggtg
     6421 ctggggttcc tggtgcaggt gttcctggag caggtgttcc tggtggcgga gttcctggcg
     6481 gtggggtgcc tggtgggggc gtgccaggcg ggggcgttcc cggtggcggt gtccccggtg
     6541 gtggggttcc agggggtggg gtaccaggcg tcggtgtacc tggtgctggt gtacccggtg
     6601 ctggagtacc gggggcaggt gtaccaggtg caggggtccc aggagctgga gttccaggtg
     6661 ctggggttcc tggtgcaggt gttcctggag caggtgttcc tggtggcgga gttcctggcg
     6721 gtggggtgcc tggtgggggc gtgccaggcg ggggcgttcc cggtggcggt gtccccggtg
     6781 gtggggttcc agggggtggg gtccctggcg tcggtgtacc tggtgctggt gtacccggtg
     6841 ctggagtacc gggggcaggt gtaccaggtg caggggtccc aggagctgga gttccaggtg
     6901 ctggggttcc tggtgcaggt gttcctggag caggtgttcc tggtggcgga gttcctggcg
     6961 gtggggtgcc tggtgggggc gtgccaggcg ggggcgttcc cggtggcggt gtccccggtg
     7021 gtggggttcc agggggtggg gtaccgggag tgggcgtccc aggtgcgggc gttcccggcg
     7081 caggtgtgcc tggagcggga gtaccaggag ccggcgtgcc aggggcgggt gtaccagggg
     7141 ccggagtgcc aggtgctgga gtaccgggcg ctggcgttcc tggtggtggc gttccgggtg
     7201 gtggagttcc cggtggtggg gttccaggtg ggggagtacc aggcggtgga gtccctggcg
     7261 gcggtgtacc aggtggtggt gtgcctggcg tcggtgtacc cggtgcaggg gttccaggag
     7321 cgggagttcc tggtgcgggt gtgccggggg ctggtgtacc tggagcaggt gttccgggag
     7381 ctggggtccc tggggcagga gtcccaggcg caggcgtccc agggggtgga gtgccgggcg
     7441 ggggtgtgcc aggtggcgga gtacctgggg gcggagtacc aggcggtggt gtgccgggcg
     7501 gcggtgtacc gggtggcggg gtacccggtg tgggagttcc gggagctggc gtcccgggtg
     7561 caggagtccc aggggctgga gtgccaggag caggcgtacc aggggcggga gtccccggag
     7621 ctggtgtccc tggtgccggg gtaccgggtg ctggcgtacc agggggcggt gtaccaggtg
     7681 gcggcgtacc tggtggtgga gttccaggcg gcggggttcc tgggggtggt gttccaggtg
     7741 ggggtgttcc cggcggcggt tctggtggc
//
