LOCUS       sequence-326342-        2660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Type IIS restriction site free elements for IBM staging 
            vectors. Staging plasmid for IBM on mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2660)
  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 2660)
  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: IBMc097
FEATURES             Location/Qualifiers
     source          1..2660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(4..98)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(119..778)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     693..712
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     940..958
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(962..1005)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1008..1029
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     complement(1104..1122)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    1620..1640
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1641..1698
                     /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(1722..1739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1893..2481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1973..1992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
       61 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
      121 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
      181 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
      241 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
      301 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgaaacgaaa aacatattct
      361 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
      421 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
      481 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
      541 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
      601 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
      661 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
      721 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
      781 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
      841 tttcattatg gtgaaagttg gaacctctta cgtgcccgat caactcgagt gccacctgac
      901 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag
      961 aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgcggccg
     1021 cttctagagg gtctcagtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1081 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1141 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     1201 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     1261 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctcct tgcaggacgg
     1321 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     1381 gcagaagaaa accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     1441 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     1501 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     1561 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtgagacct
     1621 actagtagcg gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt
     1681 tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg
     1741 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1801 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1861 aggaaccgta aaaaggccgc gttgctggcg tttttccaca ggctccgccc ccctgacgag
     1921 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1981 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2041 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2101 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2161 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2221 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2281 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2341 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2401 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2521 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2581 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2641 tggtctgaca gctcgaggct
//
