LOCUS       sequence-389373-        6710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transposon vector (Addgene #60510) for doxycycline-inducible 
            expression of a trimer with mCardinal, 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 6710)
  TITLE     Transposon vectors for proximity biotin labeling with split TurboID
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6710)
  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            "Unpublished"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/mCardinal-TurboID-GAI
FEATURES             Location/Qualifiers
     source          1..6710
                     /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             686..1417
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     CDS             1649..1675
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    2047..2168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2058..2077)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2112..2131
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             join(2891..3508,3509..3514,3515..3550,3551..3553,3554..3589,
                     3590..3592,3593..3628,3629..3634)
                     /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             3641..3697
                     /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             3698..4096
                     /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(4151..4168)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4157..4381
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4707..4724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4878..5466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4958..4977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5637..6428,6429..6497))
                     /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     6260..6279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6498..6602)
                     /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 ccaccatggt gagcaagggc gaggagctga tcaaggagaa
      721 catgcacatg aagctgtaca tggaaggcac cgtgaacaac caccacttca agtgcaccac
      781 cgaaggggag ggcaagccct acgagggcac ccagacccag aggattaagg tggtggaggg
      841 aggccccctg ccgttcgcat tcgacatcct ggccacctgc tttatgtacg ggagcaagac
      901 cttcatcaac cacacccagg gcatccccga tttctttaag cagtccttcc ctgagggctt
      961 cacatgggag agagtcacca catacgaaga cgggggcgtg cttaccgtta cccaggacac
     1021 cagcctccag gacggctgct tgatctacaa cgtcaagctc agaggggtga acttcccatc
     1081 caacggccct gtgatgcaga agaaaacact cggctgggag gccaccaccg agaccctgta
     1141 ccccgctgac ggcggcctgg aaggcagatg cgacatggcc ctgaagctcg tgggcggggg
     1201 ccacctgcac tgcaacctga agaccacata cagatccaag aaacccgcta agaacctcaa
     1261 gatgcccggc gtctactttg tggaccgcag actggaaaga atcaaggagg ccgacaatga
     1321 gacctacgtc gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact
     1381 ggggcacaaa cttaatggca tggacgagct gtacaagctc ggaggaggca gcggcggagg
     1441 atccggaggc ctggctccat atctgccacg gtgggagaag ctggataact tcatcaatag
     1501 acccgtgaag ctgatcattg gggacaaaga gattttcggg attagccggg ggattgataa
     1561 acagggagcc ctgctgctgg aacaggacgg agttatcaaa ccctggatgg gcggagaaat
     1621 cagtctgcgg tctgccgaaa agctgcagct gcctcccctg gagcgcctga ccctggacgg
     1681 cggaggaagc ggaggcggaa gtggacaaga caagaagact atgatgatga atgaagaaga
     1741 tgatggtaat ggaatggatg agcttctggc cgtgctcggt tataaggtgc gctcatccga
     1801 aatggccgac gtggcacaga agctggaaca attggaggtt atgatgtcta atgttcaaga
     1861 agatgatttg tctcagctcg ccacggagac tgttcattat aatccggcgg aactctatac
     1921 ttggctggat tctatgctga ctgatctgaa tccttgacag gcgtgggacc tctactacca
     1981 cgtgttcagg aggatcagca agcagctctg aggcctgtca ggccaagctt ccatcgatag
     2041 acatgataag atacattgat gagtttggac aaaccacaac aagaatgcag tgaaaaaaat
     2101 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2161 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     2221 aggtttttta aagcaagtaa aacctctaca aatgtggtac ctaggcgaga ccctgtctca
     2281 caaaataaag taagcccgga ctgagtgcgg aaaggcgggc ctggcgggtc tggtctcccc
     2341 atgcgggcca ccagaggccc tgcagccttc agtcgcttga aggggtaatg gcgcttccac
     2401 tcacaaacat ggcggacaga gcgtgtgaac gagatgaaca gcccctcaaa aatatggccg
     2461 ccgaggctgg acggccgtgc cccagcagca ccgcctccgc gccccacgtg atctctcgcc
     2521 gggcacagcg ctgaccgcgg aggtccaacc ggaagaatgt ccggattgga cattcggaag
     2581 agggcccgcc ttccctgggg aatctctgcg cacgcgcaga acgcttcgac caatgaaaac
     2641 acaggaagcc gtccgcgcaa ccgcgttgcg tcacttctgc cgcccctgtt tcaaggtata
     2701 tagccgtaga cggaacttcg cctttctctc ggccttagcg ccattttttt gggtgagtgt
     2761 tttttggttc ctgcgttggg attccgtgta caatccatag acatctgacc tcggcactta
     2821 gcatcatcac agcaaactaa ctgtagcctt tctctctttc cctgtagaaa cctctgcacc
     2881 tgaggccacc atgtctagac tggacaagag caaagtcata aacggcgctc tggaattact
     2941 caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc tgggagttga
     3001 gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg ccctgccaat
     3061 cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt catggcaaga
     3121 ctttctgcgg aacaacgcca agtcattccg ctgtgctctc ctctcacatc gcgacggggc
     3181 taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg aaaatcagct
     3241 cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc tgtccgccgt
     3301 gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag caaaagagga
     3361 aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa ttgagctgtt
     3421 cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca tatgtggcct
     3481 ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg attttgactt
     3541 agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc ctgctgacgc
     3601 tcttgacgat tttgaccttg acatgctccc cgggtccggc gctactaact tcagcctgct
     3661 gaagcaggct ggtgacgtcg aggagaatcc tggtcccatg gccatgcctt tgtctcaaga
     3721 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     3781 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     3841 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     3901 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     3961 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     4021 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     4081 tgtgtgggag ggctaattcg aaggcctgtc gtgaagcttg gggatcaatt ctctagagct
     4141 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4201 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4261 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4321 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4381 gcttctgagg cggaaagaac cagctgggcc tactagctac tcgggacccc ttaccgaaac
     4441 atcgccgcat tctgcagagg agtcgagtgt atgtaaactt ctgacccact gggaatgtga
     4501 tgaaagaaat aaaagctgaa atgaatcatt ctctctacta ttattctgat atttcacatt
     4561 cttaaaataa agtggtgatc ctaactgacc taagacaggg aatttttact aggattaaat
     4621 gtcaggaatt gtgaaaaagt gagtttaaat gtatttggct aaggtgtatg taaacttccg
     4681 acttcaactg tatagggatc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4741 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4801 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4861 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4921 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4981 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5041 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5101 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5221 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5281 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5341 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5401 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5461 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5521 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5581 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5641 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5701 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5761 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5821 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5881 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5941 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6001 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6061 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6601 cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag
     6661 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
//
