LOCUS       sequence-191118-        6880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Elastin Like Polypeptide: 
            [(VPGEG)-(VPGVG)4-(VPGAG)2-(VPGGG)2-(VPGEG)]6, with N-Term TagGFP2, 
            and a TEV-cleavage site, leaving an N-terminal Sortase Recognition 
            Sequence GGG and a C-terminal Cysteine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6880)
  AUTHORS   Gaub
  TITLE     Gaub Lab ELP Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6880)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET28a-HIS-TagGFP2-TEV-GGG-ELP(120nm)-Cys
FEATURES             Location/Qualifiers
     source          1..6880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(65..83)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      79..126
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      163..618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(250..269)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     460..481
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(710..1525)
                     /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(710..1525)
                     /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     1433..1452
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1517..1536
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1647..2235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2136..2155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2389..2406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2421..2560
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2486..2505)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2646..2668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2662..2853)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3662..4744)
                     /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     4706..4725
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4745..4822)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5028..5047
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5131..5150
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5131..5149
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5150..5174
                     /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             5189..5211
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5222..5239
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5954..5974
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 tgctaactcg agtaagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
       61 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      121 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
      181 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      241 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      301 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      361 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      421 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      481 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
      541 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
      601 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      661 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
      721 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
      781 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
      841 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
      901 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
      961 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     1021 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     1081 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     1141 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     1201 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     1261 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     1321 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     1381 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     1441 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
     1501 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     1561 gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1621 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1681 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1741 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     1801 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1861 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1921 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1981 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2041 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2101 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2161 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2221 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2281 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2341 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2401 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2461 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     2521 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     2581 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2641 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     2701 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     2761 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     2821 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     2881 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     2941 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     3001 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     3061 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     3121 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     3181 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     3241 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     3301 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
     3361 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     3421 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     3481 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     3541 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     3601 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     3661 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3721 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     3781 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     3841 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     3901 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     3961 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     4021 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     4081 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     4141 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     4201 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     4261 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     4321 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     4381 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     4441 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     4501 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     4561 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     4621 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     4681 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     4741 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     4801 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     4861 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     4921 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     4981 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     5041 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     5101 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
     5161 cggataacaa ttcccctgta gaaataattt tgtttaactt taagaaggag atatacatat
     5221 gcaccaccac caccaccacg gttctggtgg tgaagaactg ttcgctggta tcgttccggt
     5281 tctgatcgaa ctggacggtg acgttcacgg tcacaaattc tctgttcgtg gtgaaggtga
     5341 aggtgacgct gactacggta aactggaaat caaattcatc tgcaccaccg gtaaactgcc
     5401 ggttccgtgg ccgaccctgg ttaccaccct gtgctacggt atccagtgct tcgctcgtta
     5461 cccggaacac atgaaaatga acgacttctt caaatctgct atgccggaag gttacatcca
     5521 ggaacgtacc atccagttcc aggacgacgg taaatacaaa acccgtggtg aagttaaatt
     5581 cgaaggtgac accctggtta accgtatcga actgaaaggt aaagacttca aagaagacgg
     5641 taacatcctg ggtcacaaac tggaatactc tttcaactct cacaacgttt acatccgtcc
     5701 ggacaaagct aacaacggtc tggaagctaa cttcaaaacc cgtcacaaca tcgaaggtgg
     5761 tggtgttcag ctggctgacc actaccagac caacgttccg ctgggtgacg gtccggttct
     5821 gatcccgatc aaccactacc tgtctaccca gaccaaaatc tctaaagacc gtaacgaagc
     5881 tcgtgaccac atggttctgc tggaatcttt ctctgcttgc tgccacaccc acggtatgga
     5941 cgaactgtac cgtgaaaacc tgtacttcca ggggggcggt gtgccgggag aaggagtccc
     6001 tggtgtcggt gtcccaggcg tgggtgttcc gggtgtgggc gttccaggcg taggcgtacc
     6061 gggcgcgggt gttcctggtg ctggtgttcc gggcggcggt gttccggggg gtggcgttcc
     6121 gggtgaagga gtaccaggcg aaggcgtgcc gggtgtcggt gtcccaggcg tgggtgttcc
     6181 gggtgtgggc gttccaggcg taggcgtacc gggcgcgggt gttcctggtg ctggtgttcc
     6241 gggcggcggt gttccggggg gtggcgttcc gggtgagggt gtaccaggcg aaggggtgcc
     6301 gggtgtcggt gtcccaggcg tgggtgttcc gggtgtgggc gttccaggcg taggcgtacc
     6361 gggcgcgggt gttcctggtg ctggtgttcc gggcggcggt gttccggggg gtggcgttcc
     6421 gggtgagggt gtgccgggag aaggagtccc tggtgtcggt gtcccaggcg tgggtgttcc
     6481 gggtgtgggc gttccaggcg taggcgtacc gggcgcgggt gttcctggtg ctggtgttcc
     6541 gggcggcggt gttccggggg gtggcgttcc gggtgaagga gtaccaggcg aaggcgtgcc
     6601 gggtgtcggt gtcccaggcg tgggtgttcc gggtgtgggc gttccaggcg taggcgtacc
     6661 gggcgcgggt gttcctggtg ctggtgttcc gggcggcggt gttccggggg gtggcgttcc
     6721 gggtgagggt gtaccaggcg aaggggtgcc gggtgtcggt gtcccaggcg tgggtgttcc
     6781 gggtgtgggc gttccaggcg taggcgtacc gggcgcgggt gttcctggtg ctggtgttcc
     6841 gggcggcggt gttccggggg gtggcgttcc gggtgaaggt
//
