LOCUS       sequence-327549-        4773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Carries Golden Gate substitution insert for IBM with M86 intein. 
            PPhlF as C-lobe promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4773)
  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 4773)
  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: IBMc233
FEATURES             Location/Qualifiers
     source          1..4773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..964)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1032..1050
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1054..1097)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1100..1121
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1628..2287)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2202..2221
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3327..3346)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_RNA        3540..3589
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_feature    3816..3836
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(3842..3859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4013..4601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4093..4112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4772..4773,1..790,791..859))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      961 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1021 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     1081 tcgctaagga tgatttctgg aattcgcggc cgcttctaga ggaagacgct ggatgcatct
     1141 cgggagatag tttgatcagc ttggcgagca cagggaaaag agttcctatt aaggatttgt
     1201 taggcgaaaa agattttgaa atatgggcaa ttaatgaaca gacgatgaag ctggaatcag
     1261 ctaaagttag tcgtgtattt tgtaccggca aaaagctagt ctatactcta aaaactcgac
     1321 taggtagaac tatcaaggca acagcaaatc atagattttt aactattgat ggttggaaaa
     1381 gattagatga gctatcttta aaagagcata ttgctctacc ccgtaaacta gaataatact
     1441 agagttgaga agagaaaaga aaaccgccga tcctgtccac cgcattactg caaggtagtg
     1501 gacaagaccg gcggtcttaa gttttttggc tgaatactag agcctttggt cgaaaaaaaa
     1561 agcccgcact gtcaggtgcg ggcttttttc tgtgtttcct actagagcaa gcgagctcga
     1621 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     1681 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     1741 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     1801 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gaaacgaaaa
     1861 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     1921 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     1981 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     2041 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     2101 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     2161 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     2221 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     2281 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     2341 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccctcgg taccaaattc
     2401 cagaaaagag gcctcccgaa aggggggcct tttttcgttt tggtccaatg gcggcgcgcc
     2461 atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     2521 atgggggtga atatggcacg taccccgagc cgtagcagca ttggtagcct gcgtagtccg
     2581 catacccata aagcaattct gaccagcacc attgaaatcc tgaaagaatg tggttatagc
     2641 ggtctgagca ttgaaagcgt ggcacgtcgc gccggtgcag gcaaaccgac catttatcgt
     2701 tggtggacca acaaagcagc actgattgcc gaagtgtatg aaaatgaaat cgaacaggta
     2761 cgtaaatttc cggatttggg tagctttaaa gccgatctgg attttctgct gcataatctg
     2821 tggaaagttt ggcgtgaaac catttgtggt gaagcatttc gttgtgttat tgcagaagca
     2881 cagttggacc ctgtaaccct gacccaactg aaagatcagt ttatggaacg tcgtcgtgag
     2941 ataccgaaaa aactggttga agatgccatt agcaatggtg aactgccgaa agatatcaat
     3001 cgtgaactgc tgctggatat gatttttggt ttttgttggt atcgcctgct gaccgaacag
     3061 ttgaccgttg aacaggatat tgaagaattt accttcctgc tgattaatgg tgtttgtccg
     3121 ggtacacagt gttgataagg atcctaattg gtaacgaatc agacaattga cggctcgagg
     3181 gagtagcata gggtttgcag aatccctgct tcgtccattt gacaggcaca ttatgcatcg
     3241 atgataagct gtcaaacatg agcagatcct ctacgccgga cgcatcgtgg ccggcatcac
     3301 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     3361 ggctcgccac ttcgggctca tgagcaaata ttttatctgc caattattga aggcctccct
     3421 aacggggggc ctttttttgt ttctggtctc ccgcttaacg atcgttggct gcgacgtacg
     3481 gtggaatctg attcgttacc aattgacatg atacgaaacg taccgtatcg ttaaggtagc
     3541 tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg acgaaacagc ctctacaaat
     3601 aattttgttt aatactagag tcacacagga aagtactaga tgagctcctc tttacaattg
     3661 gcaccagaaa tagaaaagtt gcctcagagt gatatttact gggaccccat cgtttctatt
     3721 acggagactg gagtcgaaga ggtttttgat ttgactgtgc caggactacg taactttgtc
     3781 gccaatgaca tcattgtaca taactcaaga agagctacta gtagcggccg ctgcaggctt
     3841 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3901 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3961 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccaca
     4021 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4081 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4141 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4201 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4261 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4321 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4381 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4441 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4501 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4561 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4621 ctacggggtc tgacgctcag tggaaagaaa actcacgtta agggattttg gtcatgagat
     4681 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4741 aaagtatata tgagtaaact tggtctgaca gtt
//
