LOCUS       sequence-488084-        2231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Full transcription unit stuffer in an omega-level 
            cloning vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2231)
  AUTHORS   Cooke MB, Welch KT, Deus Ramirez L, Wen AX, Marciano DC,, Herman C
  TITLE     GoldenBraid2.0 E. coli: A comprehensive and characterized toolkit 
            for enterics.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2231)
  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            "Unpublished"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: pBSKo1_Stuffer1_Stuffer2
FEATURES             Location/Qualifiers
     source          1..2231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(188..776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(268..287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        891..993
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             994..1653
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1060..1079)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1766..2221
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1853..1872)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2063..2084
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggtctcagga gcgactgatg agcagactag tcgctgtcag gaggatcgct ctgccatcta
       61 gcacgctgtc atgagaccgg atcctgacac ggttatccac agaatcaggg gataacgcag
      121 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      181 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      241 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      301 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      361 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      421 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      481 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      541 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      601 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      661 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      721 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      781 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      841 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgtga tgatcggcac
      901 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
      961 ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     1021 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     1081 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     1141 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     1201 catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     1261 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     1321 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     1381 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtgtc agccaatccc
     1441 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     1501 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     1561 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     1621 cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct
     1681 taaacgcctg gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg
     1741 atgataagct gtcaaacatg atgttacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1801 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1861 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1921 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1981 tagggtgatg gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg
     2041 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2101 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2161 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2221 ttaacggatc c
//
