LOCUS       sequence-327545-        4607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Type IIS restriction site free elements for expression 
            vectors in IBM. Expression vector to generate ORF insertion library 
            in IBM on ECF20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4607)
  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 4607)
  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: IBMc326
FEATURES             Location/Qualifiers
     source          1..4607
                     /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    2755..2773
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     misc_feature    2871..2891
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2927..2958
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3469..4013
                     /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     3914..3933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4130..4147
                     /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 actcaacggc 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 gaccctgtct
     2761 cttatacaca tctattcggt ctcatctgag cggtcgtcgc aaaccgtaat actagagaac
     2821 gcatgagaaa gcccccggaa gatcaccttc cgggggcttt tttattgcgc tactagtagc
     2881 ggccgctgca ggagtcacta agggttagtt agttagatta gcagaaagtc aaaagcctcc
     2941 gaccggaggc ttttgactaa aacttccctt ggggttatca ttggggctca ctcaaaggcg
     3001 gtaatcagat aaaaaaaatc cttagctttc gctaaggatg atttctgcta gagatggaat
     3061 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3121 ctggtttatt gctgataaat ctggagccgg tgagcgtgga tctcgcggta tcattgcagc
     3181 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3241 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3301 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3361 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3421 tgagttttcg ttccactgag cgtcagaccc cttaataaga tgatcttctt gagatcgttt
     3481 tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggttttcg
     3541 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
     3601 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
     3661 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
     3721 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
     3781 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
     3841 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
     3901 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
     3961 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
     4021 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc
     4081 gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt
     4141 gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt
     4201 ttttctcctg ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc
     4261 agtatacact ccgctagcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc
     4321 aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt
     4381 tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt
     4441 tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa
     4501 gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     4561 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgtt
//
