LOCUS       sequence-428947-        3965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated minimal form of SpyCatcher with free cysteine 
            for labeling in bacteria. See also Li...Howarth, J Mol. Biol., 2014.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3965)
  AUTHORS   Zhong BL, Vachharajani VT, Dunn AR
  TITLE     Facile detection of mechanical forces across proteins in cells with 
            STReTCh.
  JOURNAL   Cell Rep Methods. 2022 Sep 2;2(9):100278. doi: 
            10.1016/j.crmeth.2022.100278. eCollection 2022 Sep 19.
  PUBMED    36160040
REFERENCE   2  (bases 1 to 3965)
  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: "Cell Rep 
            Methods."; date: "2022-09-2"; volume: "2(9):100278. doi"; pages: " 
            10.1016/j.crmeth.2022.100278. eCollection 2022 Sep 19"
COMMENT     Sequence Label: pDEST14-Cys-SpyCatcher (Min)
FEATURES             Location/Qualifiers
     source          1..3965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     173..192
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        173..191
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             223..245
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             265..282
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             304..324
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             638..655
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(708..726)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      722..769
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      806..1261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(893..912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1103..1124
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1287..1391
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1392..1460,1461..2252)
                     /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(1610..1629)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2423..3011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2912..2931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3165..3182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3197..3336
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3262..3281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3422..3444
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3438..3629)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
       61 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
      121 gcgcacccgt ggccaggacc caacgctgcc cgagatctcg atcccgcgaa attaatacga
      181 ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta actttaagaa
      241 ggagatatac atatgtcgta ctaccatcac catcaccatc acgattgcga catcccaacg
      301 accgaaaacc tgtattttca gggcgatagt gctacccata ttaaattctc aaaacgtgat
      361 gaggacggca aagagttagc tggtgcaact atggagttgc gtgattcatc tggtaaaact
      421 attagtacat ggatttcaga tggacaagtg aaagatttct acctgtatcc aggaaaatat
      481 acatttgtcg aaaccgcagc accagacggt tatgaggtag caactgctat tacctttaca
      541 gttaatgagc aaggtcaggt tactgtaaat ggctaaatgg ttgatgcttg aggatccgaa
      601 ttcgagctcc gtcgacaagc ttgcggccgc actcgagcac caccaccacc accactgaga
      661 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
      721 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
      781 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      841 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      901 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      961 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1021 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1081 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1141 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1201 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1261 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1321 attcaaatat gtatccgctc atgagacaat aaccctggta aatgcttcaa taatattgaa
     1381 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1441 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1501 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1561 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1621 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1681 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1741 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1801 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1861 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1921 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1981 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2041 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2101 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2161 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2221 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2281 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2341 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2401 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2461 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2521 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2581 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2641 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2701 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2761 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2821 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2881 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2941 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3001 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3061 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3121 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3181 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3241 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     3301 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     3361 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3421 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     3481 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     3541 tccagctcgt tgagcttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     3601 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     3661 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     3721 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     3781 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     3841 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     3901 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     3961 caggt
//
