LOCUS       sequence-332726-        4829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-fragment. This plasmid can be co-transformed with either Plasmid 
            168258: pET11a-Z-N-wtBlc or Plasmid 168473: pET11a-Z-N-DiB2 to 
            obtain wtBlc‐split-Zip or DiB2‐split-Zip protein, correspondingly .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4829)
  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 4829)
  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-Z-C-wtBlc
FEATURES             Location/Qualifiers
     source          1..4829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(388..406)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      402..449
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1051..1866)
                     /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(1051..1866)
                     /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     1774..1793
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1858..1877
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2327..2344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2461..3006)
                     /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(2541..2560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3586..4464)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4491..4775
                     /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     4698..4717
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4802..4824
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ccatggcaag cgagcagctg gaaaagaagt tacaagccct ggagaaaaaa cttgctcagc
       61 tggaatggaa aaaccaagca ttggaaaaaa aactcgcgca gacgtcgggt ggaagcggtg
      121 gtcctttcta tggcggttat aacgttattg cactcgatcg ggaataccgc catgcgctgg
      181 tttgcggccc ggaccgcgac tacctgtgga tactctcccg cacgccaacc atttctgacg
      241 aagtgaaaca ggagatgctg gcagtcgcta cccgggaagg gtttgatgtc agtaaattta
      301 tttgggtaca gcagcctggt agttagtgta caactaagga tccggctgct aacaaagccc
      361 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      421 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac
      481 aggacgggtg tggtcgccat gatggtgacc accatttgcc ccatatcacc gtcggtactg
      541 atcccgtcgt caataaaccg aaccgctaca ccctgagcat caaactcttt tatcagttgg
      601 atcatgtcgg cggtgtcgcg gccaagacgg tcgagcttct tcaccagaat gacatcacct
      661 tcctccacct tcatcctcag caaatccagc ccttcccgat ctgttgaact gccggatgcc
      721 ttgtcggtaa agatgcggtt agcttttacc cctgcatctt tgagcgctga ggtctgcctc
      781 gtgaagaagg tgttgctgac tcataccagg cctgaatcgc cccatcatcc agccagaaag
      841 tgagggagcc acggttgatg agagctttgt tgtaggtgga ccagttggtg attttgaact
      901 tttgctttgc cacggaacgg tctgcgttgt cgggaagatg cgtgatctga tccttcaact
      961 cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt caagtcagcg taatgctctg
     1021 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     1081 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1141 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1201 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1261 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg
     1321 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1381 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     1441 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1501 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     1561 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1621 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1681 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1741 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     1801 atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg
     1861 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     1921 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg
     1981 aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga
     2041 ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct
     2101 catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg cctggtatga
     2161 gtcagcaaca ccttcttcac gaggcagacc tcagcgctag cggagtgtat actggcttac
     2221 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     2281 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     2341 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     2401 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     2461 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     2521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     2581 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     2641 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     2701 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2761 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     2821 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     2881 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     2941 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     3001 tctcaagaag atcatcttat taaggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3481 attgctgcag atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
     3541 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
     3601 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
     3661 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
     3721 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
     3781 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
     3841 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
     3901 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
     3961 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
     4021 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
     4081 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     4141 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     4201 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     4261 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
     4321 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     4381 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     4441 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa
     4501 ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac
     4561 caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac
     4621 aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca
     4681 cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg
     4741 acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg ctagaaataa
     4801 ttttgtttaa ctttaagaag gagatatac
//
