LOCUS       sequence-389406-        7271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transposon vector (Addgene #60510) for doxycycline-inducible 
            expression of a trimer with human PAG1, part of TurboID (after 
            G257), and GAI (AAs 11-93) for gibberellic acid-inducible biotin 
            labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7271)
  TITLE     Transposon vectors for proximity biotin labeling with split TurboID
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7271)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSBtet-Bla/PAG1-TurboID-GAI
FEATURES             Location/Qualifiers
     source          1..7271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        295..607
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    303..321
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    339..357
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    374..392
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    410..428
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    446..464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    481..499
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    517..535
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     580..604
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      680..689
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2210..2236
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    2608..2729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2619..2638)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2673..2692
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             join(3452..4069,4070..4075,4076..4111,4112..4114,4115..4150,
                     4151..4153,4154..4189,4190..4195)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             4202..4258
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4259..4657
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAMPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(4712..4729)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4718..4942
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5268..5285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5439..6027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5519..5538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6198..6989,6990..7058))
                     /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     6821..6840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7059..7163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
       61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
      121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
      181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
      241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agacgagttt
      301 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
      361 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
      421 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
      481 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta
      541 tgtcgattag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca
      601 gatcgcctgg agccaattcc aactcttttg ccttatacct actttccgta ccacttccta
      661 ccctcgaaag gcctctgagg ccaccatggg gcccgcgggg agcctgctgg gcagcggaca
      721 gatgcagatc accctgtggg gaagtctggc tgctgtcgcc attttcttcg tcatcacctt
      781 cctcatcttc ctgtgctcta gttgtgacag ggaaaagaag ccgcgacagc atagtgggga
      841 ccatgagaac ctgatgaacg tgccttcaga caaggagatg ttcagccgtt cagttactag
      901 cctggcaaca gatgctcctg ccagcagtga gcagaatggg gcactcacca atggggacat
      961 tctttcagag gacagtactc tgacctgcat gcagcattac gaggaagtcc agacatcggc
     1021 ctcggatctg ctggattccc aggacagcac agggaaacca aaatgtcatc agagtcggga
     1081 gctgcccaga atccctcccg agagcgcagt ggataccatg ctcacggcga gaagtgtgga
     1141 cggggaccag gggctgggga tggaagggcc ctatgaagtg ctcaaggaca gctcctccca
     1201 agaaaacatg gtggaggact gcttgtatga aactgtgaaa gagatcaagg aggtggctgc
     1261 agctgcacac ctggagaaag gccacagtgg caaggcaaaa tctacttctg cctcgaaaga
     1321 gctcccaggg ccccagactg aaggcaaagc tgagtttgct gaatatgcct cggtggacag
     1381 aaacaaaaaa tgtcgtcaaa gtgttaatgt agagagtatc cttggaaatt catgtgatcc
     1441 agaagaggag gccccaccac ctgtccctgt taagcttctg gacgagaatg aaaaccttca
     1501 ggagaaggaa gggggagagg cggaagagag tgccacagac acgaccagtg aaactaacaa
     1561 gagatttagc tcattgtcat acaagtctcg ggaagaagac cccactctca cagaagaaga
     1621 gatctcagct atgtactcat cagtaaataa acctggacag ttagtgaata aatcggggca
     1681 gtcgcttaca gttccggagt ccacctacac ctccattcaa ggggacccac agaggtcacc
     1741 ctcctcctgt aatgatctct atgctactgt taaagacttc gaaaaaactc caaacagcac
     1801 acttccacca gcagggaggc ccagcgagga gccagagcct gattatgaag cgatacagac
     1861 tctcaacaga gaggaagaaa aggccaccct ggggaccaat ggccaccacg gtctcgtccc
     1921 aaaggagaac gactacgaga gcataagtga cttgcagcaa ggcagagata ttaccaggct
     1981 cggaggaggc agcggcggag gatccggagg cctggctcca tatctgccac ggtgggagaa
     2041 gctggataac ttcatcaata gacccgtgaa gctgatcatt ggggacaaag agattttcgg
     2101 gattagccgg gggattgata aacagggagc cctgctgctg gaacaggacg gagttatcaa
     2161 accctggatg ggcggagaaa tcagtctgcg gtctgccgaa aagctgcagc tgcctcccct
     2221 ggagcgcctg accctggacg gcggaggaag cggaggcgga agtggacaag acaagaagac
     2281 tatgatgatg aatgaagaag atgatggtaa tggaatggat gagcttctgg ccgtgctcgg
     2341 ttataaggtg cgctcatccg aaatggccga cgtggcacag aagctggaac aattggaggt
     2401 tatgatgtct aatgttcaag aagatgattt gtctcagctc gccacggaga ctgttcatta
     2461 taatccggcg gaactctata cttggctgga ttctatgctg actgatctga atccttgaca
     2521 ggcgtgggac ctctactacc acgtgttcag gaggatcagc aagcagctct gaggcctgtc
     2581 aggccaagct tccatcgata gacatgataa gatacattga tgagtttgga caaaccacaa
     2641 caagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2701 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2761 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2821 cctaggcgag accctgtctc acaaaataaa gtaagcccgg actgagtgcg gaaaggcggg
     2881 cctggcgggt ctggtctccc catgcgggcc accagaggcc ctgcagcctt cagtcgcttg
     2941 aaggggtaat ggcgcttcca ctcacaaaca tggcggacag agcgtgtgaa cgagatgaac
     3001 agcccctcaa aaatatggcc gccgaggctg gacggccgtg ccccagcagc accgcctccg
     3061 cgccccacgt gatctctcgc cgggcacagc gctgaccgcg gaggtccaac cggaagaatg
     3121 tccggattgg acattcggaa gagggcccgc cttccctggg gaatctctgc gcacgcgcag
     3181 aacgcttcga ccaatgaaaa cacaggaagc cgtccgcgca accgcgttgc gtcacttctg
     3241 ccgcccctgt ttcaaggtat atagccgtag acggaacttc gcctttctct cggccttagc
     3301 gccatttttt tgggtgagtg ttttttggtt cctgcgttgg gattccgtgt acaatccata
     3361 gacatctgac ctcggcactt agcatcatca cagcaaacta actgtagcct ttctctcttt
     3421 ccctgtagaa acctctgcac ctgaggccac catgtctaga ctggacaaga gcaaagtcat
     3481 aaacggcgct ctggaattac tcaatggagt cggtatcgaa ggcctgacga caaggaaact
     3541 cgctcaaaag ctgggagttg agcagcctac cctgtactgg cacgtgaaga acaagcgggc
     3601 cctgctcgat gccctgccaa tcgagatgct ggacaggcat catacccact tctgccccct
     3661 ggaaggcgag tcatggcaag actttctgcg gaacaacgcc aagtcattcc gctgtgctct
     3721 cctctcacat cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag agaaacagta
     3781 cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc
     3841 actgtacgct ctgtccgccg tgggccactt tacactgggc tgcgtattgg aggaacagga
     3901 gcatcaagta gcaaaagagg aaagagagac acctaccacc gattctatgc ccccacttct
     3961 gagacaagca attgagctgt tcgaccggca gggagccgaa cctgccttcc ttttcggcct
     4021 ggaactaatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg ggccggccga
     4081 cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg actttgacct
     4141 tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc ccgggtccgg
     4201 cgctactaac ttcagcctgc tgaagcaggc tggtgacgtc gaggagaatc ctggtcccat
     4261 ggccatgcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     4321 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     4381 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     4441 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     4501 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     4561 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     4621 tgaattgctg ccctctggtt atgtgtggga gggctaattc gaaggcctgt cgtgaagctt
     4681 ggggatcaat tctctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     4741 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4801 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4861 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4921 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggc ctactagcta
     4981 ctcgggaccc cttaccgaaa catcgccgca ttctgcagag gagtcgagtg tatgtaaact
     5041 tctgacccac tgggaatgtg atgaaagaaa taaaagctga aatgaatcat tctctctact
     5101 attattctga tatttcacat tcttaaaata aagtggtgat cctaactgac ctaagacagg
     5161 gaatttttac taggattaaa tgtcaggaat tgtgaaaaag tgagtttaaa tgtatttggc
     5221 taaggtgtat gtaaacttcc gacttcaact gtatagggat ccgcttcctc gctcactgac
     5281 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5341 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5401 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5461 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5521 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5581 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5641 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5701 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5761 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5821 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5881 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5941 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6001 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6061 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6241 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6301 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6361 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6421 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6481 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6541 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6601 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6661 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6721 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6781 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6841 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6901 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6961 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7021 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7081 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7141 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga tgcggtgtga
     7201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt
     7261 ttgttaaaat t
//
