LOCUS       sequence-332728-        4715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-fragment. This plasmid can be co-transformed with either Plasmid 
            168259: pET11a-N-wtBlc or Plasmid 168472: pET11a-N-DiB2 to obtain 
            wtBlc‐split or DiB2‐split protein, correspondingly .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4715)
  AUTHORS   Bozhanova NG, Calcutt MW, Beavers WN, Brown BP, Skaar EP, Meiler J
  TITLE     Lipocalin Blc is a potential heme-binding protein.
  JOURNAL   FEBS Lett. 2021 Jan;595(2):206-219. doi: 10.1002/1873-3468.14001. 
            Epub 2020 Dec 3.
  PUBMED    33210733
REFERENCE   2  (bases 1 to 4715)
  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"; doi: 
            "10.1002/1873-3468.14001"; journalName: "FEBS Lett"; date: 
            "2021-01"; volume: "595"; issue: "2"; pages: "206-219"
COMMENT     Sequence Label: pMRBad-C-wtBlc
FEATURES             Location/Qualifiers
     source          1..4715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(274..292)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      288..335
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(937..1752)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(937..1752)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1660..1679
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1744..1763
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2213..2230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2347..2892)
                     /direction=LEFT
                     /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     complement(2427..2446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3472..4350)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4377..4661
                     /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     4584..4603
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4688..4710
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ccatgggtcc tttctatggc ggttataacg ttattgcact cgatcgggaa taccgccatg
       61 cgctggtttg cggcccggac cgcgactacc tgtggatact ctcccgcacg ccaaccattt
      121 ctgacgaagt gaaacaggag atgctggcag tcgctacccg ggaagggttt gatgtcagta
      181 aatttatttg ggtacagcag cctggtagtt agtgtacaac taaggatccg gctgctaaca
      241 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
      301 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
      361 atccacagga cgggtgtggt cgccatgatg gtgaccacca tttgccccat atcaccgtcg
      421 gtactgatcc cgtcgtcaat aaaccgaacc gctacaccct gagcatcaaa ctcttttatc
      481 agttggatca tgtcggcggt gtcgcggcca agacggtcga gcttcttcac cagaatgaca
      541 tcaccttcct ccaccttcat cctcagcaaa tccagccctt cccgatctgt tgaactgccg
      601 gatgccttgt cggtaaagat gcggttagct tttacccctg catctttgag cgctgaggtc
      661 tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc
      721 agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt
      781 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
      841 tcaactcagc aaaagttcga tttattcaac aaagccgccg tcccgtcaag tcagcgtaat
      901 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
      961 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1021 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     1081 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1141 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1201 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1261 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     1321 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     1381 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     1441 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     1501 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     1561 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     1621 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     1681 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     1741 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     1801 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     1861 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa
     1921 cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa
     1981 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg
     2041 gtatgagtca gcaacacctt cttcacgagg cagacctcag cgctagcgga gtgtatactg
     2101 gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa
     2161 aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct tcctcgctca
     2221 ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg
     2281 gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa
     2341 gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca
     2401 gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct
     2461 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
     2521 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
     2581 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
     2641 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
     2701 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
     2761 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
     2821 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
     2881 aaacgatctc aagaagatca tcttattaag gggtctgacg ctcagtggaa cgaaaactca
     2941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3121 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3181 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3241 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3301 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3361 gttgccattg ctgcagatcg atgcataatg tgcctgtcaa atggacgaag cagggattct
     3421 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
     3481 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     3541 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     3601 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     3661 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
     3721 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     3781 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     3841 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     3901 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     3961 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     4021 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     4081 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     4141 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     4201 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     4261 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     4321 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga
     4381 aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc
     4441 gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc
     4501 catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta
     4561 tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc
     4621 tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt tttgggctag
     4681 aaataatttt gtttaacttt aagaaggaga tatac
//
