LOCUS       sequence-439439-        7581 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 7581)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 7581)
  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_miniTurbo_Nterm
FEATURES             Location/Qualifiers
     source          1..7581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(724..748)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    758..776
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    779..797
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(799..1002)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(828..848)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1003..1382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1554..1573
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1648..1752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1753..1821,1822..2613)
                     /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      2784..3372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3273..3292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3526..3543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3660..3681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3696..3701,3702..3719,3720..3726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3734..3750
                     /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     3739..3761
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3758..3774
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3758..3774
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3823..3944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3834..3853)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3888..3907
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4074..5096)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(4074..5096)
                     /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(5104..5151)
                     /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     5283..5302
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5435..5659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5648..5665
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    5707..5731
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7334..7358)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(7371..7394)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(7404..7581,1..593))
                     /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 agatatggag ccaggccttc ctgctcgaac agttccaggg ctgcccgcag ctctcggatc
       61 agcatagcgg ccagagtatt cctgtccagg ttaatccctg cttcctgcag tgtgatccag
      121 ccctgattga ccacgctttc ctccacgcgc ctcatagcca cgttaatccc ggcgccaatg
      181 acgatctgtg cagcatcgcc tgttattccg gccagctcca ccaggatgcc tgccagcttt
      241 ctatcctgca gatacaggtc attgggccat ttgactcgca ccttgtctgc tcccagcttt
      301 cgcagcgctt ctgccatgac aattccgatg accgggccca ggccgattgc tgctggtccc
      361 cgcttcaggc gccaaaacat actcaggtac aggttagctc caaaaggaga gaaccatttc
      421 ctccctctgc ttcctctccc tgcctgctga tattctgcaa tgcaagcatc cccactcttc
      481 agctcgccga ttcgatccag caggtactga ttggtggagt cgaccacagg caggactgcc
      541 acgctcccgc cgtccagctg tcccagaatc tgtttagcgt tcagcagcgg gatgctagcc
      601 atggtggcgc tagctggatc cgagctcggt accaagctta agtttaaacg ctagagtccg
      661 gaggctggat cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca
      721 ggcgatctga cggttcacta aacgagctcg tcgacgatct ctatcactga tagggagatc
      781 tctatcactg atagggagag ctctgcttat atagacctcc caccgtacac gcctaccgcc
      841 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
      901 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
      961 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     1021 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     1081 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     1141 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     1201 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     1261 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     1321 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg
     1381 tcaacgcgta tatctggccc gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag
     1441 attcttcatg caattgtcgg tcaagccttg ccttgttgta gcttaaattt tgctcgcgca
     1501 ctactcagcg acctccaaca cacaagcagg gagcagatac tggcttaact atgcggcatc
     1561 agagcagatt gtactgagag tgcaccatag gggatcggga gatctcccga tccgtcgacg
     1621 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1681 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1741 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1801 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1861 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1921 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1981 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2041 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2101 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2161 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2221 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2281 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2341 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2401 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2461 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2521 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2581 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2641 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2701 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2761 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2821 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2881 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2941 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3001 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3061 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3121 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3181 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3241 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3301 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3361 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3421 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3481 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3541 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3601 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3661 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     3721 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     3781 tacgccaagc tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt
     3841 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3901 ctattgcttt atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac
     3961 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     4021 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtagtac gtgctattcc
     4081 tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac ttctacacag
     4141 ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg tacgcccgac agtcccggct
     4201 ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg
     4261 tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc ccggagccgc
     4321 ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa
     4381 gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc cccgaacatc
     4441 gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac attgttggag
     4501 ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca aagcatcagc
     4561 tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt ttgccagtga
     4621 tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt
     4681 ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggccgc agcgatcgca
     4741 tccatggcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg caggtcttgc
     4801 aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct gaattcccca
     4861 atgtcaagca cttccggaat cgggagcgcg gccgatgcaa agtgccgata aacataacga
     4921 tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg ccctcctaca
     4981 tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg
     5041 tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttggcc
     5101 aaggaagttc ctatactttc tagagaatag gaacttcgga ataggaactt ctaggtacgt
     5161 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5221 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5281 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5341 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc
     5401 ctagagcccc agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca
     5461 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
     5521 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
     5581 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
     5641 tggcaactag aaggcacagt cgaggctgat cagcgggttt aaacgggccc tctagactcg
     5701 agtcacacca ctttgtacaa gaaagttggt caagggaagc tgtcatcatc atcttctgcc
     5761 atctccttgg caaactccaa gtcctgctgt tctcgctcac gccgttcctc tgcagactcc
     5821 aagtccttgt tgtacgattt gggaggaaac ctcatggcct tgacagacat gttgtggata
     5881 tctaggcaga aggagatgcg ctggtggaag gctagctggg gctctcgggt ggaatagatg
     5941 tcaatcatct ccttggattg gacatagccc ttctcgtggt tgatgctggc ctcaatgaca
     6001 ccatcccgga tggccttggc aacaatgaac tctgcatctt cggggctatc caactgcagc
     6061 ttctgggcga tgtcagccaa ggagattcgg gaataggaga ggctgatcat gcgtacacct
     6121 gtcttaatca cgttgtgccg cagccggata attagggtgt aggtcccatc tgcttgaaac
     6181 ttctccccaa actgatccag gacctggttg aacttggcta ggtttcctgt cctgacagct
     6241 tgagtcagaa ggaaataggg catgagtgag cgcttgaggg agggctggcg gaactgcagc
     6301 cggtcaggga tctcccccag caacagctcc accacgatga gaagcttgtg caccgtctgt
     6361 ttgaagccga cagctgtgtg ctgaggggcc ttgcgaaggg cgttggtcat cgttctccgg
     6421 gcctctgagt actccagctg gatggctttg attcgccctg tgtagtagag gtacctggcc
     6481 cactcattgt tgttggcctg ctctgggaac acagacttgg acaccagctt ctcagcctgg
     6541 tcgtacaagc tgtagtgtag gtaattccgc agcaggaggt tcaacagggt ggcctgcccg
     6601 tctgcgtcat gccgaagcgt agctgtccgg agccgagcat gcaagaagct gcgcaccaca
     6661 tccagcttgt ccaggaactc atagacccgg gcgtgataat agtaacactt tgcggctaca
     6721 aggtctaggg cccggcggtt ctgagtactg atcttctgca tcagatcatc agagatcttc
     6781 tgtgcctctt tgtagcgctt gctgttcatc atgaagatga ccacgaggag ttggagatag
     6841 gcttccactt caggcaggag gggtgtcgac gcagcttttc ccgtgcgggg acggaactgt
     6901 aaatcagcct ctgtgtccat gggctcttcc aggaagggga gcaaaaagtc tcgagtggca
     6961 ttatttgaag tgaagaagcc ctgcacagcc ttatacagaa catagtggtt gaggcggcgt
     7021 gatgtggaag gcagcatccg cagggcccgc agcacgaatc tcggctcctt gcctgaaacc
     7081 gctttctcta gctgtttcac gtgctccttg atgtcctcca aggtgactgt gtccagctct
     7141 cgctgggagt gctcagccgc tgccgccgcc gtcttgccgt ctgcctcccc cgttgacccg
     7201 ccacccgtcg ctgcctcctc tttcatctcc acatcctggg gggccggcgg cggtgggggt
     7261 tcttgttctc ctccgccggg cggcggtttc gccttgtccg cgccgcggcg ccgcgccgag
     7321 ccctcctgct tcatgccaac ttttttgtac aaacttgtgg gcgcgcctag tttatcgtca
     7381 tcgtctttgt agtcgtcctg cagcttttcg gcagaccgca gactgatttc tccgcccatc
     7441 cagggtttga taactccgtc ctgttccagc agcagggctc cctgtttatc aatcccccgg
     7501 ctaatcccga aaatctcttt gtccccaatg atcagcttca cgggtctatt gatgaagtta
     7561 tccagcttct cccaccgtga c
//
