LOCUS       sequence-439227-        7785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag-tagged biotinylation enzyme for co-expression of Psmd3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7785)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7785)
  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: PSMD3_BirA_Nterm
FEATURES             Location/Qualifiers
     source          1..7785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(232..1023,1024..1092))
                     /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     855..874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1093..1197)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1272..1291)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1463..1842
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1843..2046
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1997..2017
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2048..2066
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2069..2087
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2097..2121
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2243..2593,2594..2596,2597..3205)
                     /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             3236..3259
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3272..3296
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4899..4923)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4965..4982)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4971..5195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5328..5347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5479..5526
                     /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             5534..6556
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5534..6556
                     /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    6686..6807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6723..6742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6777..6796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6856..6872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6856..6872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6869..6891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6880..6896
                     /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(6904..6910,6911..6928,6929..6934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6949..6970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7087..7104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7258..7785,1..61))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7338..7357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
       61 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      181 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      241 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
      301 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
      361 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
      421 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
      481 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
      541 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      601 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
      661 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      721 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      781 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      841 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      901 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      961 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1021 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1081 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1141 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1201 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     1261 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     1321 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     1381 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     1441 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     1501 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     1561 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1621 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1681 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     1741 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     1801 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1861 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1921 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     1981 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2041 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
     2101 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     2161 agacaccggg accgatccag cctccggact ctagcgttta aacttaagct tggtaccgag
     2221 ctcggatcca gctagcgcca ccatgaagga caacaccgtg cccctgaagc tgatcgccct
     2281 gctggccaac ggcgagttcc acagcggcga gcagctgggc gagaccctgg gcatgagccg
     2341 ggccgccatc aacaagcaca tccagaccct gcgggactgg ggcgtggacg tgttcaccgt
     2401 gcccggcaag ggctacagcc tgcccgagcc catccagctg ctgaacgcca agcagatcct
     2461 gggccagctg gacggcggca gcgtggccgt gctgcccgtg atcgacagca ccaaccagta
     2521 cctgctggac cggatcggcg agctgaagag cggcgacgcc tgcatcgccg agtaccagca
     2581 ggccggccgg ggcggccggg gccggaagtg gttcagcccc ttcggcgcca acctgtacct
     2641 gagcatgttc tggcggctgg agcagggccc cgccgccgcc atcggcctga gcctggtgat
     2701 cggcatcgtg atggccgagg tgctgcggaa gctgggcgcc gacaaggtgc gggtgaagtg
     2761 gcccaacgac ctgtacctgc aggaccggaa gctggccggc atcctggtgg agctgaccgg
     2821 caagaccggc gacgccgccc agatcgtgat cggcgccggc atcaacatgg ccatgcggcg
     2881 ggtggaggag agcgtggtga accagggctg gatcaccctg caggaggccg gcatcaacct
     2941 ggaccggaac accctggccg ccatgctgat ccgggagctg cgggccgccc tggagctgtt
     3001 cgagcaggag ggcctggccc cctacctgag ccggtgggag aagctggaca acttcatcaa
     3061 ccggcccgtg aagctgatca tcggcgacaa ggagatcttc ggcatcagcc ggggcatcga
     3121 caagcagggc gccctgctgc tggagcagga cggcatcatc aagccctgga tgggcggcga
     3181 gatcagcctg cggagcgccg agaagggggg atctggcccc ggcggcggcg cgccagacta
     3241 caaagacgat gacgataaac taggcgcgcc cacaagtttg tacaaaaaag ttggcatgaa
     3301 gcaggagggc tcggcgcggc gccgcggcgc ggacaaggcg aaaccgccgc ccggcggagg
     3361 agaacaagaa cccccaccgc cgccggcccc ccaggatgtg gagatgaaag aggaggcagc
     3421 gacgggtggc gggtcaacgg gggaggcaga cggcaagacg gcggcggcag cggctgagca
     3481 ctcccagcga gagctggaca cagtcacctt ggaggacatc aaggagcacg tgaaacagct
     3541 agagaaagcg gtttcaggca aggagccgag attcgtgctg cgggccctgc ggatgctgcc
     3601 ttccacatca cgccgcctca accactatgt tctgtataag gctgtgcagg gcttcttcac
     3661 ttcaaataat gccactcgag actttttgct ccccttcctg gaagagccca tggacacaga
     3721 ggctgattta cagttccgtc cccgcacggg aaaagctgcg tcgacacccc tcctgcctga
     3781 agtggaagcc tatctccaac tcctcgtggt catcttcatg atgaacagca agcgctacaa
     3841 agaggcacag aagatctctg atgatctgat gcagaagatc agtactcaga accgccgggc
     3901 cctagacctt gtagccgcaa agtgttacta ttatcacgcc cgggtctatg agttcctgga
     3961 caagctggat gtggtgcgca gcttcttgca tgctcggctc cggacagcta cgcttcggca
     4021 tgacgcagac gggcaggcca ccctgttgaa cctcctgctg cggaattacc tacactacag
     4081 cttgtacgac caggctgaga agctggtgtc caagtctgtg ttcccagagc aggccaacaa
     4141 caatgagtgg gccaggtacc tctactacac agggcgaatc aaagccatcc agctggagta
     4201 ctcagaggcc cggagaacga tgaccaacgc ccttcgcaag gcccctcagc acacagctgt
     4261 cggcttcaaa cagacggtgc acaagcttct catcgtggtg gagctgttgc tgggggagat
     4321 ccctgaccgg ctgcagttcc gccagccctc cctcaagcgc tcactcatgc cctatttcct
     4381 tctgactcaa gctgtcagga caggaaacct agccaagttc aaccaggtcc tggatcagtt
     4441 tggggagaag tttcaagcag atgggaccta caccctaatt atccggctgc ggcacaacgt
     4501 gattaagaca ggtgtacgca tgatcagcct ctcctattcc cgaatctcct tggctgacat
     4561 cgcccagaag ctgcagttgg atagccccga agatgcagag ttcattgttg ccaaggccat
     4621 ccgggatggt gtcattgagg ccagcatcaa ccacgagaag ggctatgtcc aatccaagga
     4681 gatgattgac atctattcca cccgagagcc ccagctagcc ttccaccagc gcatctcctt
     4741 ctgcctagat atccacaaca tgtctgtcaa ggccatgagg tttcctccca aatcgtacaa
     4801 caaggacttg gagtctgcag aggaacggcg tgagcgagaa cagcaggact tggagtttgc
     4861 caaggagatg gcagaagatg atgatgacag cttcccttga ccaactttct tgtacaaagt
     4921 ggtgtgactc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     4981 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5041 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5101 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5161 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5221 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5461 tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg
     5521 aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     5581 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     5641 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     5701 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     5761 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     5821 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     5881 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     5941 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     6001 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     6061 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     6121 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     6181 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     6241 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     6301 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     6361 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     6421 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     6481 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     6541 gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag
     6601 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6661 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6721 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6781 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6841 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6901 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6961 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7021 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7081 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7141 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7201 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7261 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7321 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7381 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7441 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7501 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7561 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7621 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7681 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7741 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgc
//
