LOCUS       sequence-439226-        7509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag-tagged biotinylation enzyme for co-expression of Psmc3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7509)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7509)
  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: PSMC3_BirA_Nterm
FEATURES             Location/Qualifiers
     source          1..7509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     77..96
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(315..419)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(494..513)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        685..1064
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1065..1268
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1219..1239
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1270..1288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1291..1309
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1319..1343
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(1465..1815,1816..1818,1819..2427)
                     /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"
     CDS             2458..2481
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    2494..2518
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3845..3869)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3911..3928)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3917..4141
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4274..4293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4425..4472
                     /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)."
     CDS             4480..5502
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4480..5502
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5632..5753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5669..5688)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5723..5742
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5802..5818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5802..5818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5815..5837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5826..5842
                     /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)."
     promoter        complement(join(5850..5856,5857..5874,5875..5880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5895..5916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6033..6050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6204..6792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6284..6303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6963..7509,1..245,246..314))
                     /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"
ORIGIN
        1 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
       61 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      121 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      181 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      241 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      301 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      361 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      421 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
      481 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
      541 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
      601 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
      661 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
      721 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      781 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      841 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      901 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      961 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1021 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     1081 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     1141 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     1201 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1261 gcagagctct ccctatcagt gatagagatc tccctatcag tgatagagat cgtcgacgag
     1321 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1381 gaagacaccg ggaccgatcc agcctccgga ctctagcgtt taaacttaag cttggtaccg
     1441 agctcggatc cagctagcgc caccatgaag gacaacaccg tgcccctgaa gctgatcgcc
     1501 ctgctggcca acggcgagtt ccacagcggc gagcagctgg gcgagaccct gggcatgagc
     1561 cgggccgcca tcaacaagca catccagacc ctgcgggact ggggcgtgga cgtgttcacc
     1621 gtgcccggca agggctacag cctgcccgag cccatccagc tgctgaacgc caagcagatc
     1681 ctgggccagc tggacggcgg cagcgtggcc gtgctgcccg tgatcgacag caccaaccag
     1741 tacctgctgg accggatcgg cgagctgaag agcggcgacg cctgcatcgc cgagtaccag
     1801 caggccggcc ggggcggccg gggccggaag tggttcagcc ccttcggcgc caacctgtac
     1861 ctgagcatgt tctggcggct ggagcagggc cccgccgccg ccatcggcct gagcctggtg
     1921 atcggcatcg tgatggccga ggtgctgcgg aagctgggcg ccgacaaggt gcgggtgaag
     1981 tggcccaacg acctgtacct gcaggaccgg aagctggccg gcatcctggt ggagctgacc
     2041 ggcaagaccg gcgacgccgc ccagatcgtg atcggcgccg gcatcaacat ggccatgcgg
     2101 cgggtggagg agagcgtggt gaaccagggc tggatcaccc tgcaggaggc cggcatcaac
     2161 ctggaccgga acaccctggc cgccatgctg atccgggagc tgcgggccgc cctggagctg
     2221 ttcgagcagg agggcctggc cccctacctg agccggtggg agaagctgga caacttcatc
     2281 aaccggcccg tgaagctgat catcggcgac aaggagatct tcggcatcag ccggggcatc
     2341 gacaagcagg gcgccctgct gctggagcag gacggcatca tcaagccctg gatgggcggc
     2401 gagatcagcc tgcggagcgc cgagaagggg ggatctggcc ccggcggcgg cgcgccagac
     2461 tacaaagacg atgacgataa actaggcgcg cccacaagtt tgtacaaaaa agcaggctcc
     2521 accatgaatc tgctgccgaa tattgagagt ccagtgactc ggcaggagaa gatggcgacc
     2581 gtgtgggatg aggccgagca agatggaatt ggggaggagg tgctcaagat gtccacggag
     2641 gagatcatcc agcgcacacg gctgctggac agtgagatca agatcatgaa gagtgaagtg
     2701 ttgagagtca cccatgagct ccaagccatg aaggacaaga taaaagagaa cagtgagaaa
     2761 atcaaagtga acaagaccct gccgtacctt gtctccaacg tcatcgagct cctggatgtt
     2821 gatcctaatg accaagagga ggatggtgcc aatattgacc tggactccca gaggaagggc
     2881 aagtgtgctg tgatcaaaac ctctacacga cagacgtact tccttcctgt gattgggttg
     2941 gtggatgctg aaaagctaaa gccaggagac ctggtgggtg tgaacaaaga ctcctatctg
     3001 atcctggaga cgctgcccac agagtatgac tcgcgggtga aggccatgga ggtagacgag
     3061 aggcccacgg agcaatacag tgacattggg ggtttggaca agcagatcca ggagctggtg
     3121 gaggccattg tcttgccaat gaaccacaag gagaagtttg agaacttggg gatccaacct
     3181 ccaaaagggg tgctgatgta tgggccccca gggacgggga agaccctcct ggcccgggcc
     3241 tgtgccgcac agactaaggc caccttccta aagctggctg gcccccagct ggtgcagatg
     3301 ttcattggag atggtgccaa gctagtccgg gatgcctttg ccctggccaa ggagaaagcg
     3361 ccctctatca tcttcattga tgagttggat gccatcggca ccaagcgctt tgacagtgag
     3421 aaggctgggg accgggaggt gcagaggaca atgctggagc ttctgaacca gctggatggc
     3481 ttccagccca acacccaagt taaggtaatt gcagccacaa acagggtgga catcctggac
     3541 cccgccctcc tccgctcggg ccgccttgac cgcaagatag agttcccgat gcccaatgag
     3601 gaggcccggg ccagaatcat gcagatccac tcccgaaaga tgaatgtcag tcctgacgtg
     3661 aactacgagg agctggcccg ctgcacagat gacttcaatg gggcccagtg caaggctgtg
     3721 tgtgtggagg cgggcatgat cgcactgcgc aggggtgcca cggagctcac ccacgaggac
     3781 tacatggaag gcatcctgga ggtgcaggcc aagaagaaag ccaacctaca atactacgcc
     3841 taggacccag ctttcttgta caaagtggtg tgactcgagt ctagagggcc cgtttaaacc
     3901 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3961 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4021 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4081 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4141 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4201 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4261 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4321 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4381 ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga
     4441 agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg
     4501 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     4561 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
     4621 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
     4681 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc
     4741 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
     4801 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
     4861 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     4921 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     4981 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     5041 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     5101 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     5161 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     5221 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     5281 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     5341 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     5401 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     5461 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc
     5521 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5581 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5641 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5701 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5761 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5821 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5881 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5941 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6001 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6061 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6121 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6601 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6841 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6901 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6961 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7021 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7081 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7141 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7201 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7261 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7321 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7381 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7441 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7501 tctgtgact
//
