LOCUS       sequence-327544-        5110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Type IIS restriction site free elements for expressing 
            protein to be split in IBM. ECF20 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5110)
  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 5110)
  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: IBMc071
FEATURES             Location/Qualifiers
     source          1..5110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(22..27,28..44,45..50)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             98..1288
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(450..469)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      complement(1418..1447)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    1494..1515
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1516..2394)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2421..2705
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2628..2647
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    3374..3394
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3430..3461
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3972..4516
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4417..4436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4633..4650
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tactagagga gattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt
       61 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
      121 cgtcattctc ggcaccgtca ccctggacgc tgtaggcata ggcttggtta tgccggtact
      181 gccgggcctc ttgcgggata tcgtccattc cgacagtatt gccagtcact atggcgtgct
      241 gcttgcgctc tatgcgttga tgcaatttct ttgcgcaccc gttctcggag ccctgtccga
      301 ccgctttggc cgccgtccag tcctgctcgc ttcgctcctt ggagccacta tcgactacgc
      361 gatcatggcg accacacccg tcctgtggat tctctacgcc ggacgcatcg tggcgggcat
      421 cacgggtgcc acaggtgcgg ttgctggtgc ctatatcgcc gacatcaccg atggggaaga
      481 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
      541 cgtggccggg ggactgttgg gtgccatctc cttgcatgca ccattccttg cggcggcggt
      601 gctcaacggc ctcaacctcc tcctgggctg cttccttatg caggaatcgc ataagggaga
      661 gcgccgtccg atgcccttgc gtgccttcaa tccagtcagc tccttccggt gggcgcgggg
      721 catgactatc gtcgccgcac ttatgactgt tttctttatc atgcaactcg taggacaggt
      781 tccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
      841 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcacggg
      901 ccccgccacc aaacgtttcg gcgagaagca ggccattatc gcgggcatgg cggccgacgc
      961 gctgggctac gtcttgctgg cgttcgcgac gcgcggctgg atggccttcc ccattatgat
     1021 tcttctcgct tccggcggca tcggtatgcc cgcgttgcag gccatgctgt cccgccaagt
     1081 agatgacgac catcagggac agcttcaagg gtcgctcgcg gctcttacca gcctcacttc
     1141 gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     1201 gttggcatgg attgtaggtg ccgcccttta ccttgtctgc ctccccgcgt tgcgtcgcgg
     1261 tgcatggagc cgggccacct cgacctaata atactagctc cggcaaaaaa acgggcaagg
     1321 tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt tgccacctga
     1381 cgtctaagaa aaggaatatt cagcaatttg cccgtgccga agaaaggccc acccgtgaag
     1441 gtgagccagt gagttgattg ctacgtaatt agttagttag cccttagtga ctcgaattcg
     1501 cggccgcttc tagagttatg acaacttgac ggctacatca ttcacttttt cttcacaacc
     1561 ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag
     1621 ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa
     1681 aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa
     1741 ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct
     1801 ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac
     1861 ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa
     1921 ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt
     1981 aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa
     2041 ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg
     2101 aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat
     2161 ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt
     2221 tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg
     2281 tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag
     2341 atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt
     2401 catactcccg ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca
     2461 ctgcgtcttt tactggcact tctcgctaac caaaccggta accccgctta ttaaaagcat
     2521 tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca
     2581 cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt
     2641 tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc
     2701 tccattacta gagtcacaca ggactactag atgaatgaaa ccgatcctga tctggaactg
     2761 ctgaaacgta ttggtaataa tgatgcacag gccgttaaag aaatggttac ccgtaaactg
     2821 cctcgtctgc tggcactggc aagtcgcctg ctgggtgatg cagatgaagc acgtgatatt
     2881 gcacaagaaa gttttctgcg catttggaaa caggcagcaa gctggcgtag cgaacaggca
     2941 cgttttgata cctggctgca tcgtgttgca ctgaatctgt gttatgatcg tctgcgtcgt
     3001 cgtaaagaac atgtgccggt tgatagcgaa catgcctgtg aagcactgga tacccgtccg
     3061 gcaccggatg aacagctgga agcaagcgca cagagccgtc gtatggcaca ggcactggat
     3121 cagctgccgg atcgtcagcg tgaagcaatt gttctgcagt attatcaaga actgagcaat
     3181 accgaagcag cagcactgat gcaaattagc gttgaagccc tggaaagcct gctgagccgt
     3241 gcacgtcgta atctgcgtag ccatctggcc gaagcaccgg gtgcagatct gagcggtcgt
     3301 cgcaaaccgt aatactagag aacgcatgag aaagcccccg gaagatcacc ttccgggggc
     3361 ttttttattg cgctactagt agcggccgct gcaggagtca ctaagggtta gttagttaga
     3421 ttagcagaaa gtcaaaagcc tccgaccgga ggcttttgac taaaacttcc cttggggtta
     3481 tcattggggc tcactcaaag gcggtaatca gataaaaaaa atccttagct ttcgctaagg
     3541 atgatttctg ctagagatgg aatagactgg atggaggcgg ataaagttgc aggaccactt
     3601 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3661 ggatctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3841 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3901 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccttaata
     3961 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     4021 accgccttgc agggcggttt tcgaaggttc tctgagctac caactctttg aaccgaggta
     4081 actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc
     4141 atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt
     4201 gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac
     4261 tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt
     4321 gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa
     4381 ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag
     4441 tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg
     4501 gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc
     4561 ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga
     4621 acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc
     4681 tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct
     4741 catcagtgcc aacatagtaa gccagtatac actccgctag cgctgaggtc tgcctcgtga
     4801 agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc agaaagtgag
     4861 ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt tgaacttttg
     4921 ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct tcaactcagc
     4981 aaaagttcga tttattcaac aaagccacgt tgtgtctcaa aatctctgat gttacattgc
     5041 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     5101 aaggggtgtt
//
