LOCUS       sequence-439440-        7305 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 7305)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7305)
  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_miniTurbo_Nterm
FEATURES             Location/Qualifiers
     source          1..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      461..470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(586..610)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    620..638
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    641..659
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(661..864)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(690..710)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        865..1244
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1416..1435
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1510..1614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1615..1683,1684..2475)
                     /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      2646..3234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3135..3154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3388..3405
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3522..3543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3558..3563,3564..3581,3582..3588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3596..3612
                     /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     3601..3623
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3620..3636
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3620..3636
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3685..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3696..3715)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3750..3769
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3936..4958)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(3936..4958)
                     /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
                     "
     protein_bind    complement(4966..5013)
                     /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     5145..5164
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5297..5521
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5510..5527
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    5569..5593
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6920..6944)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(6957..6980)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(6990..7305,1..455))
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
ORIGIN
        1 tcctccacgc gcctcatagc cacgttaatc ccggcgccaa tgacgatctg tgcagcatcg
       61 cctgttattc cggccagctc caccaggatg cctgccagct ttctatcctg cagatacagg
      121 tcattgggcc atttgactcg caccttgtct gctcccagct ttcgcagcgc ttctgccatg
      181 acaattccga tgaccgggcc caggccgatt gctgctggtc cccgcttcag gcgccaaaac
      241 atactcaggt acaggttagc tccaaaagga gagaaccatt tcctccctct gcttcctctc
      301 cctgcctgct gatattctgc aatgcaagca tccccactct tcagctcgcc gattcgatcc
      361 agcaggtact gattggtgga gtcgaccaca ggcaggactg ccacgctccc gccgtccagc
      421 tgtcccagaa tctgtttagc gttcagcagc gggatgctag ccatggtggc gctagctgga
      481 tccgagctcg gtaccaagct taagtttaaa cgctagagtc cggaggctgg atcggtcccg
      541 gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac
      601 taaacgagct cgtcgacgat ctctatcact gatagggaga tctctatcac tgatagggag
      661 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
      721 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
      781 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
      841 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
      901 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
      961 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     1021 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     1081 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     1141 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     1201 ccccgtaatt gattactatt aataactagt caataatcaa tgtcaacgcg tatatctggc
     1261 ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc agattcttca tgcaattgtc
     1321 ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa
     1381 cacacaagca gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     1441 agtgcaccat aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg
     1501 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1561 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1621 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1681 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1741 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1801 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1861 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1921 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1981 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2041 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2101 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2161 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2221 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2281 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2341 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2401 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2461 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2521 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2581 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2641 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2701 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2761 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2821 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2881 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2941 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3001 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3061 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3121 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3181 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3241 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3301 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3361 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3421 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3481 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     3541 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     3601 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     3661 gaggtcgacg gtatacagac atgataagat acattgatga gtttggacaa accacaacta
     3721 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3781 ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg
     3841 cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag
     3901 aaggcggcgg tggaatcgaa atctcgtagt acgtgctatt cctttgccct cggacgagtg
     3961 ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc
     4021 gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg
     4081 tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag
     4141 agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc
     4201 ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag
     4261 aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg
     4321 accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg
     4381 aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg
     4441 acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca
     4501 atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg
     4561 ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc
     4621 ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca
     4681 cggcgggaga tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga
     4741 atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg
     4801 gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga
     4861 gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga
     4921 aacttctcga cagacgtcgc ggtgagttca ggctttttgg ccaaggaagt tcctatactt
     4981 tctagagaat aggaacttcg gaataggaac ttctaggtac gtgaaccatc accctaatca
     5041 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5101 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5161 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5221 gccgcgctta atgcgccgct acagggcgcg tggggatacc ccctagagcc ccagctggtt
     5281 ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc
     5341 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     5401 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     5461 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     5521 gtcgaggctg atcagcgggt ttaaacgggc cctctagact cgagtcacac cactttgtac
     5581 aagaaagctg ggtcctaggc gtagtattgt aggttggctt tcttcttggc ctgcacctcc
     5641 aggatgcctt ccatgtagtc ctcgtgggtg agctccgtgg cacccctgcg cagtgcgatc
     5701 atgcccgcct ccacacacac agccttgcac tgggccccat tgaagtcatc tgtgcagcgg
     5761 gccagctcct cgtagttcac gtcaggactg acattcatct ttcgggagtg gatctgcatg
     5821 attctggccc gggcctcctc attgggcatc gggaactcta tcttgcggtc aaggcggccc
     5881 gagcggagga gggcggggtc caggatgtcc accctgtttg tggctgcaat taccttaact
     5941 tgggtgttgg gctggaagcc atccagctgg ttcagaagct ccagcattgt cctctgcacc
     6001 tcccggtccc cagccttctc actgtcaaag cgcttggtgc cgatggcatc caactcatca
     6061 atgaagatga tagagggcgc tttctccttg gccagggcaa aggcatcccg gactagcttg
     6121 gcaccatctc caatgaacat ctgcaccagc tgggggccag ccagctttag gaaggtggcc
     6181 ttagtctgtg cggcacaggc ccgggccagg agggtcttcc ccgtccctgg gggcccatac
     6241 atcagcaccc cttttggagg ttggatcccc aagttctcaa acttctcctt gtggttcatt
     6301 ggcaagacaa tggcctccac cagctcctgg atctgcttgt ccaaaccccc aatgtcactg
     6361 tattgctccg tgggcctctc gtctacctcc atggccttca cccgcgagtc atactctgtg
     6421 ggcagcgtct ccaggatcag ataggagtct ttgttcacac ccaccaggtc tcctggcttt
     6481 agcttttcag catccaccaa cccaatcaca ggaaggaagt acgtctgtcg tgtagaggtt
     6541 ttgatcacag cacacttgcc cttcctctgg gagtccaggt caatattggc accatcctcc
     6601 tcttggtcat taggatcaac atccaggagc tcgatgacgt tggagacaag gtacggcagg
     6661 gtcttgttca ctttgatttt ctcactgttc tcttttatct tgtccttcat ggcttggagc
     6721 tcatgggtga ctctcaacac ttcactcttc atgatcttga tctcactgtc cagcagccgt
     6781 gtgcgctgga tgatctcctc cgtggacatc ttgagcacct cctccccaat tccatcttgc
     6841 tcggcctcat cccacacggt cgccatcttc tcctgccgag tcactggact ctcaatattc
     6901 ggcagcagat tcatggtgga gcctgctttt ttgtacaaac ttgtgggcgc gcctagttta
     6961 tcgtcatcgt ctttgtagtc gtcctgcagc ttttcggcag accgcagact gatttctccg
     7021 cccatccagg gtttgataac tccgtcctgt tccagcagca gggctccctg tttatcaatc
     7081 ccccggctaa tcccgaaaat ctctttgtcc ccaatgatca gcttcacggg tctattgatg
     7141 aagttatcca gcttctccca ccgtgacaga tatggagcca ggccttcctg ctcgaacagt
     7201 tccagggctg cccgcagctc tcggatcagc atagcggcca gagtattcct gtccaggtta
     7261 atccctgctt cctgcagtgt gatccagccc tgattgacca cgctt
//
