LOCUS       sequence-180326-        6562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CBM-Xmod-DocIII handle with N-Terminal Sortase Recognition Sequence 
            GGG for posttranslational coupling to a C-Terminal LPETGG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6562)
  AUTHORS   Ott W, Jobst MA, Bauer MS, Durner E, Milles LF, Nash MA, Gaub HE
  TITLE     Elastin-like Polypeptide Linkers for Single-Molecule Force 
            Spectroscopy.
  JOURNAL   ACS Nano. 2017 Jun 12. doi: 10.1021/acsnano.7b02694.
  PUBMED    28591514
REFERENCE   2  (bases 1 to 6562)
  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: "ACS Nano. 
            2017 Jun 12. doi: 10.1021/acsnano.7b02694."
COMMENT     Sequence Label: pET28a-GGG-HIS-CBM-Xmod-DocIII
FEATURES             Location/Qualifiers
     source          1..6562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     940..959
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(979..1056)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1262..1281
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1365..1384
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1365..1383
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1384..1408
                     /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             1423..1445
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1465..1482
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2861..2879)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2875..2922
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2959..3414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3046..3065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3256..3277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3506..4321)
                     /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(3506..4321)
                     /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     4229..4248
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4313..4332
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4443..5031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4932..4951
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5185..5202
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5217..5356
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5282..5301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5442..5464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5458..5649)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6458..6562,1..978))
                     /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"
ORIGIN
        1 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
       61 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
      121 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
      181 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
      241 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
      301 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
      361 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
      421 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
      481 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
      541 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
      601 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
      661 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
      721 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
      781 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
      841 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
      901 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
      961 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     1021 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     1081 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     1141 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     1201 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     1261 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     1321 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
     1381 aggggaattg tgagcggata acaattcccc tgtagaaata attttgttta actttaagaa
     1441 ggagatatac atatgggggg cggtcaccac caccaccacc acggaatggc caatacaccg
     1501 gtatcaggca atttgaaggt tgaattctac aacagcaatc cttcagatac tactaactca
     1561 atcaatcctc agttcaaggt tactaatacc ggaagcagtg caattgattt gtccaaactc
     1621 acattgagat attattatac agtagacggt cagaaagatc agaccttctg gagtgaccat
     1681 gctgcaataa tcggcagtaa cggcagctac aacggaatta cttcaaatgt aaaaggaaca
     1741 tttgtaaaaa tgagttcctc aacaaataac gcagacacct accttgaaat aagctttaca
     1801 ggcggaactc ttgaaccggg tgcacatgtt cagatacaag gtagatttgc aaagaatgac
     1861 tggagtaact atacacagtc aaatgactac tcattcaagt ctgcttcaca gtttgttgaa
     1921 tgggatcagg taacagcata cttgaacggt gttcttgtat ggggtaaaga acccggtggc
     1981 agtgtagtac catcaacaca gcctgtaaca acaccacctg caacaacaaa accacctgca
     2041 acaacaatac cgccgtcaga tgatccgaat gccgtggtac caaacactgt tacatcagct
     2101 gtaaagacac agtatgttga gatcgaatct gttgacggct tctacttcaa cacagaagac
     2161 aagttcgata cagctcagat caagaaggct gttctccaca cagtttataa cgaaggctac
     2221 acaggtgatg acggcgtggc tgtagttctc agagaatatg agagcgagcc tgttgatatc
     2281 acagctgagt taacattcgg tgatgcaaca cctgctaaca catacaaggc tgttgaaaac
     2341 aagttcgatt acgagatccc tgtttactac aacaacgcta cactcaagga tgctgagggc
     2401 aacgatgcta cagtaactgt ttacatcggt ctcaagggcg atacagacct taacaacatc
     2461 gttgacggtc gtgacgctac agcaacactt acttactacg ctgcaacatc tacagacggc
     2521 aaggatgcta caacagttgc actttcacca agcacactgg ttggcggcaa ccctgagtca
     2581 gtatatgatg acttctctgc attccttagc gatgttaagg tagacgctgg taaggaactc
     2641 acaagattcg ctaagaaggc tgagagactt atcgacggta gagatgcttc aagcatactc
     2701 acattctaca ctaagtcttc tgtagatcag tacaaggata tggctgctaa cgagccaaac
     2761 aagctttggg atatcgttac aggcgatgct gaagaagaat aactcgagta agatccggct
     2821 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     2881 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     2941 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3001 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3061 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3121 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3181 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3241 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3301 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3361 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     3421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3481 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     3541 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     3601 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     3661 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     3721 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     3781 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     3841 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     3901 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     3961 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     4021 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     4081 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     4141 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     4201 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     4261 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     4321 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc
     4381 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4441 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4501 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4561 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4621 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4681 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4741 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4801 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4861 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4921 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4981 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5041 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5101 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5161 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     5221 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     5281 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     5341 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5401 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5461 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5521 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5581 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5641 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     5701 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     5761 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     5821 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     5881 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     5941 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     6001 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     6061 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
     6121 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     6181 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     6241 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     6301 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     6361 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     6421 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6481 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6541 tattgggcgc cagggtggtt tt
//
