LOCUS       sequence-221161-        4802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell expression vector for BirA Biotin Ligase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4802)
  AUTHORS   Moon JJ, Pepper M
  TITLE     Generation of Allergen-Specific Tetramers for a Murine Model of 
            Airway Inflammation.
  JOURNAL   Methods Mol Biol. 2018;1799:165-181. doi: 
            10.1007/978-1-4939-7896-0_14.
  PUBMED    29956152
REFERENCE   2  (bases 1 to 4802)
  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: "Methods Mol
            Biol."; date: "2018"; volume: "1799:165-181. doi"; pages: " 
            10.1007/978-1-4939-7896-0_14"
COMMENT     Sequence Label: pR BirA
FEATURES             Location/Qualifiers
     source          1..4802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..431
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             439..1404
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             439..1401
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(2186..2203)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2186..2202)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2195..2217)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2411..2430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2530..2552
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2590..2608)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2676..2780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2781..2849,2850..3641)
                     /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"
     rep_origin      3812..4400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4301..4320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4554..4571
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4688..4709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4724..4729,4730..4747,4748..4754)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4762..4778
                     /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     4767..4789
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4786..4802
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4786..4802
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgattaccg ttgcaggaca ggatgtggtg cccgatgtga ctagctcttt gctgcaggcc
       61 gtcctatcct ctggttccga taagagaccc agaactccgg ccccccaccg cccaccgcca
      121 cccccataca tatgtggtac gcaagtaaga gtgcctgcgc atgccccatg tgccccacca
      181 agagctttgc atcccataca agtccccaaa gtggagaacc gaaccaattc ttcgcgggca
      241 gaacaaaagc ttctgcacac gtctccactc gaatttggag ccggccggcg tgtgcaaaag
      301 aggtgaatcg aacgaaagac ccgtgtgtaa agccgcgttt ccaaaatgta taaaaccgag
      361 agcatctggc caatgtgcat cagttgtggt cagcagcaaa atcaagtgaa tcatctcagt
      421 gcaactaaag gggaattcat gaaggataac accgtgccac tgaaattgat tgccctgtta
      481 gcgaacggtg aatttcactc tggcgagcag ttgggtgaaa cgctgggaat gagccgggcg
      541 gctattaata aacacattca gacactgcgt gactggggcg ttgatgtctt taccgttccg
      601 ggtaaaggat acagcctgcc tgagcctatc cagttactta atgctaaaca gatattgggt
      661 cagctggatg gcggtagtgt agccgtgctg ccagtgattg actccacgaa tcagtacctt
      721 cttgatcgta tcggagagct taaatcgggc gatgcttgca ttgcagaata ccagcaggct
      781 ggccgtggtc gccggggtcg gaaatggttt tcgccttttg gcgcaaactt atatttgtcg
      841 atgttctggc gtctggaaca aggcccggcg gcggcgattg gtttaagtct ggttatcggt
      901 atcgtgatgg cggaagtatt acgcaagctg ggtgcagata aagttcgtgt taaatggcct
      961 aatgacctct atctgcagga tcgcaagctg gcaggcattc tggtggagct gactggcaaa
     1021 actggcgatg cggcgcaaat agtcattgga gccgggatca acatggcaat gcgccgtgtt
     1081 gaagagagtg tcgttaatca ggggtggatc acgctgcagg aagcggggat caatctcgat
     1141 cgtaatacgt tggcggccat gctaatacgt gaattacgtg ctgcgttgga actcttcgaa
     1201 caagaaggat tggcacctta tctgtcgcgc tgggaaaagc tggataattt tattaatcgc
     1261 ccagtgaaac ttatcattgg tgataaagaa atatttggca tttcacgcgg aatagacaaa
     1321 cagggggctt tattacttga gcaggatgga ataataaaac cctggatggg cggtgaaata
     1381 tccctgcgta gtgcagaaaa ataaggatcc tctagagtcg acctgcaggc atgcaagcta
     1441 ttcgatgcac actcacattc ttctcctaat acgataataa aactttccat gaaaaatatg
     1501 gaaaaatata tgaaaattga gaaatccaaa aaactgataa acgctctact taattaaaat
     1561 agataaatgg gagcggcagg aatggcggag catggccaag ttcctccgcc aatcagtcgt
     1621 aaaacagaag tcgtggaaag cggatagaaa gaatgttcga tttgacgggc aagcatgtct
     1681 gctatgtggc ggattgcgga ggaattgcac tggagaccag caaggttctc atgaccaaga
     1741 atatagcggt gagtgagcgg gaagctcggt ttctgtccag atcgaactca aaacttgtcc
     1801 agccagtcgc tgtcgaaact aattaagtta atgagttttt catgttagtt tcgcgctgag
     1861 caacaattaa gtttatgttt cagttcggct tagatttcgc tgaaggactt gccactttca
     1921 atcaatactt tagaacaaaa tcaaaactca ttctaatagc ttggtgttca tctttttttt
     1981 taatgataag cattttgtcg tttatacttt ttatatttcg atattaaacc acctatgaag
     2041 tctattttaa tcgccagata agcaatatat tgtgtaaata tttgtattct ttatcaggaa
     2101 attcagggag acggggaagt tactatctac taaaagccaa acaatttctt acagttttac
     2161 tctctctact ctagagtagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     2221 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2281 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2341 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     2401 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     2461 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2521 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2581 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2641 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2701 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2761 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2821 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2881 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2941 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3001 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3061 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3121 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3181 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3241 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3301 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3361 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3421 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3481 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3541 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3601 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3661 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3721 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3781 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3841 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3901 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3961 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4021 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4081 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4141 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4201 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4261 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4321 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4381 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4441 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4501 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4561 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4621 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     4681 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     4741 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     4801 ac
//
