LOCUS       sequence-327546-        3600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Carries engineered transposon for IBM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3600)
  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 3600)
  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: IBMc090
FEATURES             Location/Qualifiers
     source          1..3600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      183..771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     672..691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     925..942
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1132..1151)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1143..1958
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQERLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1216..1235)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2131..2427)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     2277..2299
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2291..2308
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2292..2308
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2318..2337
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2318..2336
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2384..2404)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2384..2402)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2423..2439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2423..2439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2436..2458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2447..2463
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2471..2477,2478..2495,2496..2501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2516..2537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2646..2664)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             2826..3485
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2892..2911)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3506..3600
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 ccaagcctcg agctgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
       61 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      121 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      181 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      241 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      301 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      361 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      421 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      481 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      541 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      601 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      661 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      721 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctgtggaa aaacgccagc
      781 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      841 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      901 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcctg cataacgcga
      961 agatctggag cgtcttccga aaaacgcgaa agcgtttcac gataaatgcg aaaactgtct
     1021 ggatctgaat aattaattaa attgtgtctc aaaatctctg atgttacatt gcacaagata
     1081 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     1141 ttatgagcca tattcaacgg gaaacgtctt gttcgaggcc gcgattaaat tccaacatgg
     1201 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     1261 tctatcggtt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     1321 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     1381 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     1441 cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     1501 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     1561 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     1621 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     1681 aagaaatgca taagctgttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     1741 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     1801 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     1861 ctccttcatt acaggaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     1921 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattacta gagttgagaa
     1981 gagaaaagaa aaccgccgat cctgtccacc gcattactgc aaggtagtgg acaagaccgg
     2041 cggtcttaag ttttttggct gaatactaga gcctttggtc gaaaaaaaaa gcccgcactg
     2101 tcaggtgcgg gcttttttct gtgtttccta ctagccgcta cagggcgcgt cccattcgcc
     2161 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     2221 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     2281 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     2341 attgggtacc gggccccccc tcgaggtcct ccagcttttg ttccctttag tgagggttaa
     2401 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2461 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2521 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccaccgg ttgacccata
     2581 atgttttcgc atttatcgtg aaacgctttc gcgtttttcg tgctcttctt cagatctgaa
     2641 attctgcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2701 gacgtcaggt ggcactcgag ttgatcgggc acgtaagagg ttccaacttt caccataatg
     2761 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     2821 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2881 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2941 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     3001 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     3061 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     3121 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     3181 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     3241 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     3301 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     3361 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     3421 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     3481 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa
     3541 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
//
