LOCUS       sequence-437133-        7763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enterokinase-responsive elastin-like polypeptide (ELP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7763)
  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 7763)
  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-EK-cleavableELP
FEATURES             Location/Qualifiers
     source          1..7763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             919..936
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(989..1007)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1003..1050
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1087..1542
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1174..1193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1384..1405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1568..1672
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1673..1741,1742..2533)
                     /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(1891..1910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2704..3292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3193..3212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3446..3463
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3478..3617
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3543..3562)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3703..3725
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3719..3910)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4719..5801)
                     /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     5763..5782
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5802..5879)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6085..6104
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6188..6207
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6188..6206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6207..6231
                     /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             6246..6268
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7746..7760
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 ggcggctccg tgccaggcgt cggtgtgcct ggagtcgggg tgccaggggt aggagtccct
       61 ggcgtaggag tgccaggagt gggtgttccg ggtgtaggag ttcctggagt gggggttccc
      121 ggtgttggcg taccgggcgt tggagtacct ggtgtaggtg tgcccggtgt aggcgtgcct
      181 ggcgttggtg ttccaggagt tggggtcccc ggagttggcg ttccaggtgt cggggttccg
      241 ggggttggag tgcctggtgt gggcgtacct ggagtaggag tacccggtgt gggagtgccg
      301 ggagttggag tcccaggggt cggcgttcct ggggtgggag tcccgggggt gggcgttccc
      361 ggagttgggg tgcccggagt aggcgtacca ggcgtgggag taccgggtgt gggggtccca
      421 ggtgtagggg tacccggcgt gggtgtgccg ggtgtcggag ttcccggagt cggagtcccc
      481 ggggtgggtg taccgggggt cggggtacca ggggtggggg tgccgggggt aggcgttccg
      541 ggcgtagggg ttcctggtgt cggtgttccc ggcgtaggtg ttcctggcgt gggggtaccg
      601 ggagtgggcg tcccgggtgt tggagttccg ggagtaggtg tacccggagt gggagttcca
      661 ggcgttgggg ttccaggggt tggcgtccca ggagtaggtg tacctggggt aggtgtccct
      721 ggtgttggtg tacctggcgt cggagtgccc ggggttgggg tgcctggggt tggtgtgcca
      781 ggtgttgggg tacctggggt cggagtacca ggagtcggtg taccaggtgt gggtgtccca
      841 ggcgtaggcg tccctggagt tggtgtcccc ggtgtcggcg tacccggggt cggtgtcccg
      901 ggagtcggct ggctcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
      961 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     1021 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
     1081 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1141 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     1201 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     1261 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     1321 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     1381 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1441 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1501 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     1561 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1621 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1681 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1741 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1801 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1861 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1921 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1981 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2041 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2101 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2161 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     2221 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2281 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2341 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggct ctcgcggtat
     2401 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2461 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2521 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2581 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2641 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2701 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2761 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2821 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2881 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2941 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3001 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3061 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3121 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3181 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3241 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3301 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3361 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3421 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3481 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     3541 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     3601 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3661 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3721 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     3781 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     3841 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3901 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     3961 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     4021 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     4081 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     4141 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     4201 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     4261 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     4321 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     4381 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     4441 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     4501 tcgcgcgcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     4561 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4621 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     4681 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4741 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4801 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     4861 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     4921 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     4981 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     5041 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     5101 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     5161 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     5221 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     5281 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     5341 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     5401 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     5461 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     5521 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     5581 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     5641 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     5701 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     5761 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     5821 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     5881 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     5941 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     6001 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     6061 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     6121 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     6181 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     6241 ataattttgt ttaactttaa gaaggagata tacatatggg tggttgggga ccagtgcctg
     6301 gcgtcggtgt acctggtgct ggtgtacccg gtgctggagt accgggggca ggtgtaccag
     6361 gtgcaggggt cccaggagct ggagttccag gtgctggggt tcctggtgca ggtgttcctg
     6421 gagcaggtgt tcctggtggc ggagttcctg gcggtggggt gcctggtggg ggcgtgccag
     6481 gcgggggcgt tcccggtggc ggtgtccccg gtggtggggt tccagggggt ggggtaccag
     6541 gcgtcggtgt acctggtgct ggtgtacccg gtgctggagt accgggggca ggtgtaccag
     6601 gtgcaggggt cccaggagct ggagttccag gtgctggggt tcctggtgca ggtgttcctg
     6661 gagcaggtgt tcctggtggc ggagttcctg gcggtggggt gcctggtggg ggcgtgccag
     6721 gcgggggcgt tcccggtggc ggtgtccccg gtggtggggt tccagggggt ggggtccctg
     6781 gcgtcggtgt acctggtgct ggtgtacccg gtgctggagt accgggggca ggtgtaccag
     6841 gtgcaggggt cccaggagct ggagttccag gtgctggggt tcctggtgca ggtgttcctg
     6901 gagcaggtgt tcctggtggc ggagttcctg gcggtggggt gcctggtggg ggcgtgccag
     6961 gcgggggcgt tcccggtggc ggtgtccccg gtggtggggt tccagggggt ggggtaccgg
     7021 gagtgggcgt cccaggtgcg ggcgttcccg gcgcaggtgt gcctggagcg ggagtaccag
     7081 gagccggcgt gccaggggcg ggtgtaccag gggccggagt gccaggtgct ggagtaccgg
     7141 gcgctggcgt tcctggtggt ggcgttccgg gtggtggagt tcccggtggt ggggttccag
     7201 gtgggggagt accaggcggt ggagtccctg gcggcggtgt accaggtggt ggtgtgcctg
     7261 gcgtcggtgt acccggtgca ggggttccag gagcgggagt tcctggtgcg ggtgtgccgg
     7321 gggctggtgt acctggagca ggtgttccgg gagctggggt ccctggggca ggagtcccag
     7381 gcgcaggcgt cccagggggt ggagtgccgg gcgggggtgt gccaggtggc ggagtacctg
     7441 ggggcggagt accaggcggt ggtgtgccgg gcggcggtgt accgggtggc ggggtacccg
     7501 gtgtgggagt tccgggagct ggcgtcccgg gtgcaggagt cccaggggct ggagtgccag
     7561 gagcaggcgt accaggggcg ggagtccccg gagctggtgt ccctggtgcc ggggtaccgg
     7621 gtgctggcgt accagggggc ggtgtaccag gtggcggcgt acctggtggt ggagttccag
     7681 gcggcggggt tcctgggggt ggtgttccag gtgggggtgt tcccggcggc ggttctggtg
     7741 gcggtgatga cgatgataaa ggt
//
