LOCUS       sequence-326355-        2633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Type IIS restriction site free elements for IBM staging 
            vectors. Additional BsaI site to facilitate size selection. Staging 
            plasmid for IBM on ECF20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2633)
  AUTHORS   Ho TYH, Shao A, Lu Z, Savilahti H, Menolascina F, Wang L, Dalchau N,
            Wang B
  TITLE     A systematic approach to inserting split inteins for Boolean logic 
            gate engineering and basal activity reduction.
  JOURNAL   Nat Commun. 2021 Apr 13;12(1):2200. doi: 10.1038/s41467-021-22404-9.
  PUBMED    33850130
REFERENCE   2  (bases 1 to 2633)
  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: "Nat 
            Commun."; date: "2021-04-13"; volume: "12(1):2200. doi"; pages: " 
            10.1038/s41467-021-22404-9"
COMMENT     Sequence Label: IBMc367
FEATURES             Location/Qualifiers
     source          1..2633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(106..200)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(221..880)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     795..814
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1042..1060
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1064..1107)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1110..1131
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_feature    1694..1714
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1715..1772
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1796..1813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1967..2555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2047..2066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
       61 taaagtatat atgagtaaac ttggtctcga cagctcgagg cttggattct caccaataaa
      121 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
      181 tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact
      241 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
      301 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
      361 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
      421 tgaaactcac ccagggattg gctgaaacga aaaacatatt ctcaataaac cctttaggga
      481 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
      541 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
      601 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
      661 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
      721 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
      781 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
      841 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
      901 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
      961 tggaacctct tacgtgcccg atcaactcga gtgccacctg acgtctaaga aaccattatt
     1021 atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa
     1081 tccttagctt tcgctaagga tgatttctgg aattcgcggc cgcttctaga gggtctcaat
     1141 cctgatctgg aactgctgaa acgtattggt aataatgatg cacaggccgt taaagaaatg
     1201 gttacccgta aactgcctcg tctgctggca ctggcaagtc gcctgctggg tgatgcagat
     1261 gaagcacgtg atattgcaca agaaagtttt ctgcgcattt ggaaacaggc agcaagctgg
     1321 cgtagcgaac aggcacgttt tgatacctgg ctgcatcgtg ttgcactgaa tctgtgttat
     1381 gatcgtctgc gtcgtcgtaa agaacatgtg ccggttgata gcgaacatgc ctgtgaagca
     1441 ctggataccc gtccggcacc ggatgaacag ctggaagcaa gcgcacagag ccgtcgtatg
     1501 gcacaggcac tggatcagct gccggatcgt cagcgtgaag caattgttct gcagtattat
     1561 caagaactga gcaataccga agcagcagca ctgatgcaaa ttagcgttga agccctggaa
     1621 agcctgctga gccgtgcacg tcgtaatctg cgtagccatc tggccgaagc accgggtgca
     1681 gatctgtgag acctactagt agcggccgct gcagtccggc aaaaaagggc aaggtgtcac
     1741 caccctgccc tttttcttta aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc
     1801 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1861 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cacaggctcc
     1981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2401 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2521 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aga
//
