LOCUS       sequence-326343-        2594 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 TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2594)
  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 2594)
  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: IBMc228
FEATURES             Location/Qualifiers
     source          1..2594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..660)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     575..594
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     822..840
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(844..887)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    890..911
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     complement(936..955)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    1435..1455
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1456..1513
                     /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(1537..1554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1708..2296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1788..1807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2480..2574)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
       61 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
      121 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
      181 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgaaacga aaaacatatt
      241 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
      301 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
      361 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
      421 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
      481 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
      541 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
      601 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
      661 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
      721 tatttcatta tggtgaaagt tggaacctct tacgtgcccg atcaactcga gtgccacctg
      781 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
      841 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattcgcggc
      901 cgcttctaga gggtctcagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa
      961 gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga
     1021 cgccttagcc attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga
     1081 aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca
     1141 tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct
     1201 cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc
     1261 actcagcgct gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt
     1321 cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc
     1381 tatcgaatta tttgatcacc aaggtgcaga gccagccttc ttattcgtga gacctactag
     1441 tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt
     1501 aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact cgctgcgctc
     1561 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1621 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1681 ccgtaaaaag gccgcgttgc tggcgttttt ccacaggctc cgcccccctg acgagcatca
     1741 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1801 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1861 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1921 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1981 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2041 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2101 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2161 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2221 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2281 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2341 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2401 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2461 tcgacagctc gaggcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     2521 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     2581 agctcgatat caaa
//
