LOCUS       sequence-488085-        2228 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 2228)
  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  JOURNAL   Unpublished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 2228)
  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            "Unpublished"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: pBSKo2_Stuffer2_Stuffer1
FEATURES             Location/Qualifiers
     source          1..2228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(187..775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(267..286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        890..992
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             993..1652
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1059..1078)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1765..2220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1852..1871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2062..2083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggtctcagtc aggaggatcg ctctgccatc tagcacgctg tcaggagcga ctgatgagca
       61 gactagtcgc ttgagacctg cagtgacacg gttatccaca gaatcagggg ataacgcagg
      121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      601 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgtgat gatcggcacg
      901 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
      961 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     1021 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     1081 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     1141 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     1201 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     1261 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     1321 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     1381 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtgtca gccaatccct
     1441 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     1501 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     1561 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     1621 agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt
     1681 aaacgcctgg ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga
     1741 tgataagctg tcaaacatga tgttacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1801 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1861 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1921 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1981 agggtgatgg ttcacgtagt gggccagcgc cctgatagac ggtttttcgc cctttgacgt
     2041 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2101 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2161 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2221 taactgca
//
