LOCUS       sequence-333251-        4490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  wild‐type E. coli lipocalin Blc without signal peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4490)
  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 4490)
  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: pBAD-wtBlc
FEATURES             Location/Qualifiers
     source          1..4490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      28..55
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        74..165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(166..234,235..1026)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(384..403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1197..1785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1686..1705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1939..1956
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1971..2111
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2037..2056)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2197..2219
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2375..3253)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3280..3564
                     /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     3487..3506
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3610..3627
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4170..4187)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4340..4426
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
       61 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      121 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      181 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      241 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      301 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      361 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
      421 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      481 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      541 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      601 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      661 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      721 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      781 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      841 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      901 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      961 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1021 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1081 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1321 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1441 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1501 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1561 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1621 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1681 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1741 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1801 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1861 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1921 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     1981 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     2041 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     2101 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2161 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2221 tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc
     2281 ggcatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
     2341 ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat
     2401 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
     2461 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
     2521 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
     2581 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
     2641 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
     2701 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
     2761 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
     2821 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
     2881 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
     2941 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
     3001 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
     3061 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
     3121 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
     3181 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
     3241 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
     3301 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
     3361 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
     3421 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
     3481 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
     3541 cgcaactctc tactgtttct ccatacccgt tttttgggct aacaggagga attaaccatg
     3601 gggggttctc atcatcatca tcatcatctc gaatcaacca gcttgtacaa gaaatccagc
     3661 tccacgccgc cgcgtggcgt gaccgtagta aataatttcg acgccaaacg ttatctcggt
     3721 acctggtatg agattgcccg ttttgatcac cgctttgaac gtggactgga aaaagtcacc
     3781 gcaacataca gcctgcgtga tgacggcggc ctgaatgtca ttaataaagg ctataaccct
     3841 gacagaggaa tgtggcagca gagtgaaggg aaagcgtact ttaccggcgc accaactcgc
     3901 gctgcgctga aagtgtcatt ctttggtcct ttctatggcg gttataacgt tattgcactc
     3961 gatcgggaat accgccatgc gctggtttgc ggcccggacc gcgactacct gtggatactc
     4021 tcccgcacgc caaccatttc tgacgaagtg aaacaggaga tgctggcagt cgcgacccgg
     4081 gaagggtttg atgtcagtaa atttatttgg gtacagcagc ctggtagtta gaagcttggc
     4141 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     4201 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     4261 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     4321 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4381 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     4441 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
//
