LOCUS       sequence-232146-        5918 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses BirA and eCPX-AP(K10).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5918)
  AUTHORS   Granhoj J, Dimke H, Svenningsen P
  TITLE     A bacterial display system for effective selection of protein-biotin
            ligase BirA variants with novel peptide specificity.
  JOURNAL   Sci Rep. 2019 Mar 11;9(1):4118. doi: 10.1038/s41598-019-40984-x.
  PUBMED    30858523
REFERENCE   2  (bases 1 to 5918)
  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: "Sci Rep."; 
            date: "2019-03-11"; volume: "9(1):4118. doi"; pages: " 
            10.1038/s41598-019-40984-x"
COMMENT     Sequence Label: pBAD-BirA-eCPX-AP(K10)
FEATURES             Location/Qualifiers
     source          1..5918
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             312..334
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             346..1308
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             346..1308
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1309..1326
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1384..1401)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1554..1640
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1732..1759
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1779..1870
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1871..1939,1940..2731)
                     /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(2089..2108)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2902..3490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3391..3410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3549..3596)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3592..3610
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    4396..4420
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(4420..4439)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4421..4439)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4578..4595
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4610..4750
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4676..4695)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4836..4858
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5014..5892)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagaaataa ttttgtttaa ctttaagaag gagatataca tacccatgaa ggataacacc
      361 gtgccactga aattgattgc cctgttagcg aacggtgaat ttcactctgg cgagcagttg
      421 ggtgaaacgc tgggaatgag ccgggcggct attaataaac acattcagac actgcgtgac
      481 tggggcgttg atgtctttac cgttccgggt aaaggataca gcctgcctga gcctatccag
      541 ttacttaatg ctaaacagat attgggtcag ctggatggcg gtagtgtagc cgtgctgcca
      601 gtgattgact ccacgaatca gtaccttctt gatcgtatcg gagagcttaa atcgggcgat
      661 gcttgcattg cagaatacca gcaggctggc cgtggtcgcc ggggtcggaa atggttttcg
      721 ccttttggcg caaacttata tttgtcgatg ttctggcgtc tggaacaagg cccggcggcg
      781 gcgattggtt taagtctggt tatcggtatc gtgatggcgg aagtattacg caagctgggt
      841 gcagataaag ttcgtgttaa atggcctaat gacctctatc tgcaggatcg caagctggca
      901 ggcattctgg tggagctgac tggcaaaact ggcgatgcgg cgcaaatagt cattggagcc
      961 gggatcaaca tggcaatgcg ccgtgttgaa gagagtgtcg ttaatcaggg gtggatcacg
     1021 ctgcaggaag cggggatcaa tctcgatcgt aatacgttgg cggccatgct aatacgtgaa
     1081 ttacgtgctg cgttggaact cttcgaacaa gaaggattgg caccttatct gtcgcgctgg
     1141 gaaaagctgg ataattttat taatcgccca gtgaaactta tcattggtga taaagaaata
     1201 tttggcattt cacgcggaat agacaaacag ggggctttat tacttgagca ggatggaata
     1261 ataaaaccct ggatgggcgg tgaaatatcc ctgcgtagtg cagaaaaaca tcatcatcat
     1321 catcattgag agtttaaacg gtctccagct tggctgtttt ggcggatgag agaagatttt
     1381 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     1441 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     1501 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     1561 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1621 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     1681 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     1741 tcctgacgga tggccttttt gcgtttctac aaactctttt tgtttatttt tctaaataca
     1801 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1861 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1921 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1981 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2041 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2101 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2161 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2221 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2281 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2341 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2401 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2461 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2521 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2581 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2641 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2701 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2761 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2821 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2881 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2941 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3001 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3061 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3121 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3181 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3241 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3301 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3361 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3421 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3481 cctatggaaa aacgccagca acgcggcctt tttacggttc ctgaaccgga tatagttcct
     3541 cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
     3601 tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatca
     3661 gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg aattcgttta
     3721 aacattactt aagctcgtgc cactcgattg cctgtgcttc aaaaatatca ttcagaccgc
     3781 ggccgcctgc acgacgtgca ccgcttgcgg tacggctttt ttcggtatag gtaaagctac
     3841 caataacacc cagcggacta ttatcttctt cgtagcgata tttcagatta aagccaccca
     3901 ttttattcat ctgaccctgt gcatcactct gtgcataacc accggtaacg gtgctggttg
     3961 cacgacggct tttgctacca aaacgataac caacgctcag aatccaggtg ccaacatcaa
     4021 cgctacgaat acggctctgt tcatagctga aatccagtgc aacattttcc atcggattaa
     4081 actgcagacc tgcaccatag ctaaaaccat aatcgctggt atcatgttta taggtcggat
     4141 attcggtggt ctgaaattta ccataaccaa cacccacaac accataaatg cttgcccaat
     4201 cattaatacg atatgccgga ccggctgtaa tgccgtaata ctgatttttg ttataatcac
     4261 cactctgacc gctctgacca ccggcaacgc tggtgcctgc ggtaaatgcc agaactgctg
     4321 ccagtgcgct cagacatgcg atttttttca tggcgatcgc tccttactaa agttaaagaa
     4381 aattattact agtggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa
     4441 tttcgcggga tcgagatccc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4501 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4561 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4621 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     4681 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4741 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4801 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4861 ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga aggcgaagcg
     4921 gcatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc
     4981 cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt
     5041 cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa
     5101 tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg
     5161 catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct
     5221 taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca
     5281 aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta
     5341 ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc
     5401 catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc
     5461 ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc
     5521 ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg
     5581 ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg
     5641 acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac
     5701 aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata
     5761 acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg
     5821 cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg
     5881 cgcttcagcc atacttttca tactcccgcc attcagag
//
