LOCUS       sequence-326357-        4272 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 TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4272)
  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 4272)
  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: IBMc227
FEATURES             Location/Qualifiers
     source          1..4272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(174..242,243..1034)
                     /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"
     primer_bind     complement(392..411)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(1161..1190)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    1237..1258
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1259..2137)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2164..2448
                     /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     2371..2390
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    2699..2719
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2755..2786
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3297..3841
                     /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     3742..3761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3958..3975
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cgatttattc aacaaagcca
       61 cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
      121 aataaaactg tctgcttaca taaacagtaa tacaaggggt gtttactaga ggtatgagta
      181 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      241 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      301 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      361 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      421 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      481 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      541 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      601 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      661 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      721 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      781 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      841 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga tctcgcggta
      901 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      961 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1021 ttaagcattg gtaatactag ctccggcaaa aaaacgggca aggtgtcacc accctgccct
     1081 ttttctttaa aaccgaaaag attacttcgc gtttgccacc tgacgtctaa gaaaaggaat
     1141 attcagcaat ttgcccgtgc cgaagaaagg cccacccgtg aaggtgagcc agtgagttga
     1201 ttgctacgta attagttagt tagcccttag tgactcgaat tcgcggccgc ttctagagtt
     1261 atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg
     1321 ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa
     1381 cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc
     1441 tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat
     1501 gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc
     1561 tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg
     1621 gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta
     1681 tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca
     1741 ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata
     1801 ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt
     1861 gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca
     1921 gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac
     1981 cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat
     2041 cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt
     2101 atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca
     2161 gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc
     2221 acttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga
     2281 ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca
     2341 ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag
     2401 cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatta ctagagggaa
     2461 gagagtcaat tcagggtggt gaatatgtcc agattagata aaagtaaagt gattaacagc
     2521 gcattagagc tgcttaatga ggtctgagac cacggccaca tcgtggaggt ctcattcggc
     2581 cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtcctgatac
     2641 tagagaacgc atgagaaagc ccccggaaga tcaccttccg ggggcttttt tattgcgcta
     2701 ctagtagcgg ccgctgcagg agtcactaag ggttagttag ttagattagc agaaagtcaa
     2761 aagcctccga ccggaggctt ttgactaaaa cttcccttgg ggttatcatt ggggctcact
     2821 caaaggcggt aatcagataa aaaaaatcct tagctttcgc taaggatgat ttctgctaga
     2881 gatggaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2941 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggatc tcgcggtatc
     3001 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3061 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3121 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3181 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3241 ccttaacgtg agttttcgtt ccactgagcg tcagacccct taataagatg atcttcttga
     3301 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc
     3361 ggttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag
     3421 cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta
     3481 actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg
     3541 ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg
     3601 tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg
     3661 agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga
     3721 gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3781 ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa
     3841 aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg
     3901 aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc
     3961 gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt
     4021 gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat
     4081 agtaagccag tatacactcc gctagcgctg aggtctgcct cgtgaagaag gtgttgctga
     4141 ctcataccag gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttgat
     4201 gagagctttg ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg
     4261 gtctgcgttg tc
//
