LOCUS       sequence-183394-        1713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An ORF part-containing vector harbouring the SpyTag sequence with 4 
            bp overhangs specific for the 'upstream SpyPart' position within an 
            ORF assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1713)
  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 1713)
  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-B-SpyTag-C
FEATURES             Location/Qualifiers
     source          1..1713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      37..71
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(95..133)
                     /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(250..838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(330..349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1020..1679
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1086..1105)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
       61 gcgtttttta ttggtctggt cggtctcatg agcccttcgt cggtttataa gcatcaacca
      121 ttacaatatg agcgctttga gaccccgact acggttatcc acagaatcag gggataacgc
      181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      661 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      901 gattttggtc atgattgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa
      961 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     1021 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     1081 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     1141 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     1201 acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg
     1261 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1321 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1381 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1441 atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     1501 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     1561 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     1621 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat
     1681 ttgatatcga gctcgcttgg actcctgttg ata
//
