LOCUS       sequence-439230-        7973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag-tagged biotinylation enzyme for co-expression of Lmod1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7973)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7973)
  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"
COMMENT     Sequence Label: Lmod1-BirA
FEATURES             Location/Qualifiers
     source          1..7973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(25..633,634..636,637..987))
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     protein_bind    998..1022
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2844..2868)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2962..2986)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2996..3014
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3017..3035
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3037..3240)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3066..3086)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3241..3620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3792..3811
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3886..3990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3991..4059,4060..4851)
                     /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"
     primer_bind     complement(4209..4228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5022..5610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5511..5530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5764..5781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5898..5919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5934..5939,5940..5957,5958..5964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5972..5988
                     /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     5977..5999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5996..6012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5996..6012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6061..6182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6072..6091)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6126..6145
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6312..7334)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(7342..7389)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     7521..7540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    7673..7897
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7886..7903
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(7944..7967)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 gccgccgccg gggccagatc cccccttctc ggcgctccgc aggctgatct cgccgcccat
       61 ccagggcttg atgatgccgt cctgctccag cagcagggcg ccctgcttgt cgatgccccg
      121 gctgatgccg aagatctcct tgtcgccgat gatcagcttc acgggccggt tgatgaagtt
      181 gtccagcttc tcccaccggc tcaggtaggg ggccaggccc tcctgctcga acagctccag
      241 ggcggcccgc agctcccgga tcagcatggc ggccagggtg ttccggtcca ggttgatgcc
      301 ggcctcctgc agggtgatcc agccctggtt caccacgctc tcctccaccc gccgcatggc
      361 catgttgatg ccggcgccga tcacgatctg ggcggcgtcg ccggtcttgc cggtcagctc
      421 caccaggatg ccggccagct tccggtcctg caggtacagg tcgttgggcc acttcacccg
      481 caccttgtcg gcgcccagct tccgcagcac ctcggccatc acgatgccga tcaccaggct
      541 caggccgatg gcggcggcgg ggccctgctc cagccgccag aacatgctca ggtacaggtt
      601 ggcgccgaag gggctgaacc acttccggcc ccggccgccc cggccggcct gctggtactc
      661 ggcgatgcag gcgtcgccgc tcttcagctc gccgatccgg tccagcaggt actggttggt
      721 gctgtcgatc acgggcagca cggccacgct gccgccgtcc agctggccca ggatctgctt
      781 ggcgttcagc agctggatgg gctcgggcag gctgtagccc ttgccgggca cggtgaacac
      841 gtccacgccc cagtcccgca gggtctggat gtgcttgttg atggcggccc ggctcatgcc
      901 cagggtctcg cccagctgct cgccgctgtg gaactcgccg ttggccagca gggcgatcag
      961 cttcaggggc acggtgttgt ccttcattct agaatcaacc actttgtaca agaaagctgg
     1021 gtcggcgcgc ccaccctttt gaagtagctt gggcacctct accttcttca gctgcttaag
     1081 gttgctagaa cggatggcag ccaggagctg gtcacgcgag ttcttctcct gggcagggag
     1141 cactttgtct cccatcttcc tctgggtagc tggtgagagt gagttcttta ggttctccat
     1201 gataaggggc ggggccaggg gagggggagg gggaggcggg gctgctggaa cacctgcttt
     1261 cttgggtgag ttctttggag caggctttgg agatggctgg ggtgaaggtt tgggggagcc
     1321 cttggccaga gccccgacct tgggtacctc cagccggtcc ttcttctctc cactggcttc
     1381 ctgggcctgc ttttgctctt gcagccgttt ctgtcgctgc ttgtccatgt ttcggctgag
     1441 cagattggtg acagtcatcc ggggtcctgc cagctcaaaa tggtagccca gcttgagcaa
     1501 ggtggtgttc tccttgagca gcttggcgat ctccatctca gtcttgcccc cacagatgtg
     1561 cctctggttg tgaaagcgca gctctgtcag agtgttgttc tggaggaggg cccggaagat
     1621 ggccaggatg cctttgccag tgatgtgatt ggagtccagg ttgaggctgg tgatggtctt
     1681 attggccttt aacatgatag caattgcaaa ggccacatgg tcgtcagccc gagtattggc
     1741 caaggcaaac accttcacca cagtgttgaa ctccagggcc tctgtgaagc ggaccaggat
     1801 ctcgttggtg atgcagtctg agttgttgac gttcacctct gtcatctctg ggtcgttgtt
     1861 cttcactttc tccagaggtt catcaaatat gctgggagct gcttcctctt ctgctctggc
     1921 ttgcccatca gagggcttgt tggggcaact gggcgcctgt ttctccgggg ctttggtctt
     1981 gctctcttcc ctgacttctt tgtcaggttg gttttcttcc ttcttaactt tttcatcctc
     2041 ttttctccag tctctgcccc cagaacctat gccctcatcc tctggtttct tgttctcctt
     2101 gcttgactct ttcagcctcc cgtcctcccg tctcccgact gtgtttctgc tctctgctgt
     2161 cagtttcacc tcctctttct tgctgggctc cttcacctct tctttcttct tgtctttgtc
     2221 tctgctcaga cctgcgttct tgacactccc tgtcttttct tcctccttcc ttgcagcggc
     2281 tgccctctct tctctgccat ccttcccaga ctccttcctg tccaccgcag ccctgaccct
     2341 gcctttatca atacctctga tgaccttctc ctccttgggc ttctcacccc ctctgccatc
     2401 ggcctcctct cgatccctgg agaagctttt ctttaaaacg cccttcttcg gctccttgcc
     2461 caggtctgag tcctgtctgg ggcctggggc ctttctgctg gcatcgctgt ccttctcctc
     2521 tccattcttg gcatctgtct tcctccccac ttgcttgctt tcatccacag acatctctct
     2581 ctggatgagt ttcttgctct ccttttcaca gaagttgagc atggcttccc ggttgaatga
     2641 gcccgagggc tgtttgtccg tttgattcct ctgccgcagc cccaccggga tgctcccgtc
     2701 tgggtccacc acatctagtt ccttctccag ctcctccatc tcttcagggg agagggtgga
     2761 cagcaggcta tcgatgtcgg ggtcttcact cacctgccgc cggtacttag ctactttgga
     2821 catggtgaag ggggcggccg cggagcctgc ttttttgtac aaacttgtct aaagcttaag
     2881 tttaaacgct agagtccgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca
     2941 gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctcgtc gacgatctct
     3001 atcactgata gggagatctc tatcactgat agggagagct ctgcttatat agacctccca
     3061 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     3121 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     3181 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     3241 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     3301 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     3361 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     3421 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     3481 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     3541 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     3601 actagtcaat aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa
     3661 acgcctaacc ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc
     3721 ttaaattttg ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg
     3781 gcttaactat gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga
     3841 tctcccgatc cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3901 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3961 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4021 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4081 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4141 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4201 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4261 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4321 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4381 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4441 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4501 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4561 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4621 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4681 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4741 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4801 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4861 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4921 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4981 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5041 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5101 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5161 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5221 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5281 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5341 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5401 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5461 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5521 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5581 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5641 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5701 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5761 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5821 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5881 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5941 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6001 aacagctatg accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga
     6061 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     6121 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     6181 ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     6241 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     6301 cgtagtacgt gctattcctt tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg
     6361 gcgagtactt ctacacagcc atcggtccag acggccgcgc ttctgcgggc gatttgtgta
     6421 cgcccgacag tcccggctcc ggatcggacg attgcgtcgc atcgaccctg cgcccaagct
     6481 gcatcatcga aattgccgtc aaccaagctc tgatagagtt ggtcaagacc aatgcggagc
     6541 atatacgccc ggagccgcgg cgatcctgca agctccggat gcctccgctc gaagtagcgc
     6601 gtctgctgct ccatacaagc caaccacggc ctccagaaga agatgttggc gacctcgtat
     6661 tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc
     6721 gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc
     6781 tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc
     6841 atcacagttt gccagtgata cacatgggga tcagcaatcg cgcatatgaa atcacgccat
     6901 gtagtgtatt gaccgattcc ttgcggtccg aatgggccga acccgctcgt ctggctaaga
     6961 tcggccgcag cgatcgcatc catggcctcc gcgaccggct gcagaacagc gggcagttcg
     7021 gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc gggagatgca ataggtcagg
     7081 ctctcgctga actccccaat gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag
     7141 tgccgataaa cataacgatc tttgtagaaa ccatcggcgc agctatttac ccgcaggaca
     7201 tatccacgcc ctcctacatc gaagctgaaa gcacgagatt cttcgccctc cgagagctgc
     7261 atcaggtcgg agacgctgtc gaacttttcg atcagaaact tctcgacaga cgtcgcggtg
     7321 agttcaggct ttttggccaa ggaagttcct atactttcta gagaatagga acttcggaat
     7381 aggaacttct aggtacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     7441 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     7501 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     7561 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     7621 ggcgcgtggg gataccccct agagccccag ctggttcttt ccgcctcaga agccatagag
     7681 cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc
     7741 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca
     7801 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag
     7861 gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttaa
     7921 acgggccctc tagactcgag ttatttatcg tcatcgtctt tgtagtctgg cgc
//
