LOCUS       sequence-314117-        4567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Carries Golden Gate substitution insert for IBM with M86 intein. 
            Plux2 as C-lobe promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4567)
  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 4567)
  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: IBMc050
FEATURES             Location/Qualifiers
     source          1..4567
                     /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"
     promoter        complement(2288..2390)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     RBS             3246..3257
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3269..3303)
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     terminator      3311..3338
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3610..3630
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(3636..3653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3807..4395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3887..3906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4566..4567,1..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 gttcgatata 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 gtgcctacta gagggaccaa
     2401 aacgaaaaaa gacgcttttc agcgtcttat tgttcgtctt tggtaccgag ctctagtagt
     2461 gatctacact agcactatca gtgttattaa tttttaaagt atgggcaatc aattgctcct
     2521 gttaaaattg ctttagaaat actttggcag cggtttgttg tattgagttt catttgcgca
     2581 ttggttaaat ggaaagtgac agtacgctca ctgcaaccta atatttttga aatatcccaa
     2641 gagctttttc cttcgcatgc ccacgctaaa cattcttttt ctcttttggt taaatcgttg
     2701 tttgatttat tatttgctat atttattttt cgataattat caactagaga aggaacaatt
     2761 aatggtatgt tcatacacgc atgtaaaaat aaactatcta tatagttgtc tttttctgaa
     2821 tgtgcaaaac taagcattcc gaagccattg ttagccgtat gaatagggaa actaaaccca
     2881 gtgataagac ctgatgtttt cgcttcttta attacatttg gagatttttt atttacagca
     2941 ttgttttcaa atatattcca attaattggt gaatgattgg agttagaata atctactata
     3001 ggatcatatt ttattaaatt agcgtcatca taatattgcc tccatttttt agggtaatta
     3061 tctaggattg aaatatcaga tttaaccata gaatgaggat aaatgatcgc gagtaaataa
     3121 tattcacaat gtaccatttt agtcatatca gataagcatt gattaatatc attattgctt
     3181 ctacaagctt taattttatt aattattctg tatgtgtcgt cggcatttat gtttttcatc
     3241 tagtatttct cctctttaat ctctagtagc tagcacaatc cctaggactg agctagctgt
     3301 caatcacact ggctcacctt cgggtgggcc tttctgcgtt tatatactag acacctgtag
     3361 gatcgtacag gtttacgcaa gaaaatggtt tgttactttc gaataatact agagtcacac
     3421 aggaaagtac tagatgagct cctctttaca attggcacca gaaatagaaa agttgcctca
     3481 gagtgatatt tactgggacc ccatcgtttc tattacggag actggagtcg aagaggtttt
     3541 tgatttgact gtgccaggac tacgtaactt tgtcgccaat gacatcattg tacataactc
     3601 aagaagagct actagtagcg gccgctgcag gcttcctcgc tcactgactc gctgcgctcg
     3661 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3721 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3781 cgtaaaaagg ccgcgttgct ggcgtttttc cacaggctcc gcccccctga cgagcatcac
     3841 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3901 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3961 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4021 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4081 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4141 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4201 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4261 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4321 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4381 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4441 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4501 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4561 gacagtt
//
