LOCUS       sequence-183404-        1746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An ORF part-containing vector harbouring the SpyTag sequence with a 
            His6 tag at its C-terminus and 4 bp overhangs specific for the 
            'downstream SpyPart' position within an ORF assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1746)
  AUTHORS   Gilbert C, Howarth M, Harwood CR, Ellis T
  TITLE     Extracellular Self-Assembly of Functional and Tunable Protein 
            Conjugates from Bacillus subtilis.
  JOURNAL   ACS Synth Biol. 2017 Mar 7. doi: 10.1021/acssynbio.6b00292.
  PUBMED    28230977
REFERENCE   2  (bases 1 to 1746)
  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 Synth 
            Biol. 2017 Mar 7. doi: 10.1021/acssynbio.6b00292."
COMMENT     Sequence Label: pCG-D-SpyTag-His6-E
FEATURES             Location/Qualifiers
     source          1..1746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      477..511
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(542..559)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(566..604)
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     rep_origin      complement(723..1311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(803..822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1493..1746,1..406)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1559..1578)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
       61 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
      121 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
      181 ttcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
      241 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
      301 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
      361 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg atatcgagct
      421 cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
      481 gaacgctcgg ttgccgccgg gcgtttttta ttggtctggt cggtctcaga acttagctgc
      541 cgtgatgatg atgatgatgg cttccctttg taggtttgta cgcgtctacc ataacgatgt
      601 gggcgctgcc tgagaccccg actacggtta tccacagaat caggggataa cgcaggaaag
      661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      901 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1141 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1261 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1321 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1381 gtcatgattg atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact
     1441 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
     1501 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     1561 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     1621 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     1681 tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt
     1741 gatatg
//
