LOCUS       sequence-389395-        6104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transposon vector (Addgene #60510) for doxycycline-inducible 
            expression of EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6104)
  TITLE     Transposon vectors for proximity biotin labeling with split TurboID
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6104)
  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            "Unpublished"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/GFP
FEATURES             Location/Qualifiers
     source          1..6104
                     /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             join(686..688,689..691,692..1405)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(731..752)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(992..1011)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1339..1360
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1441..1562
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1452..1471)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1506..1525
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             join(2285..2902,2903..2908,2909..2944,2945..2947,2948..2983,
                     2984..2986,2987..3022,3023..3028)
                     /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             3035..3091
                     /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             3092..3490
                     /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(3545..3562)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3551..3775
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4101..4118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4272..4860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4352..4371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5031..5822,5823..5891))
                     /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     5654..5673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5892..5996)
                     /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 gaggagctgt tcaccggggt
      721 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      781 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      841 caagctgccc gtgccctggc ccaccctcgt gaccaccttc acctacggcg tgcagtgctt
      901 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      961 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1021 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1081 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaaggtcta
     1141 tatcaccgcc gacaagcaga agaacggcat caaggtgaac ttcaagaccc gccacaacat
     1201 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1261 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     1321 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1381 cggcatggac gagctgtaca agtaaggcct gtcaggccaa gcttccatcg atagacatga
     1441 taagatacat tgatgagttt ggacaaacca caacaagaat gcagtgaaaa aaatgcttta
     1501 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1561 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     1621 tttaaagcaa gtaaaacctc tacaaatgtg gtacctaggc gagaccctgt ctcacaaaat
     1681 aaagtaagcc cggactgagt gcggaaaggc gggcctggcg ggtctggtct ccccatgcgg
     1741 gccaccagag gccctgcagc cttcagtcgc ttgaaggggt aatggcgctt ccactcacaa
     1801 acatggcgga cagagcgtgt gaacgagatg aacagcccct caaaaatatg gccgccgagg
     1861 ctggacggcc gtgccccagc agcaccgcct ccgcgcccca cgtgatctct cgccgggcac
     1921 agcgctgacc gcggaggtcc aaccggaaga atgtccggat tggacattcg gaagagggcc
     1981 cgccttccct ggggaatctc tgcgcacgcg cagaacgctt cgaccaatga aaacacagga
     2041 agccgtccgc gcaaccgcgt tgcgtcactt ctgccgcccc tgtttcaagg tatatagccg
     2101 tagacggaac ttcgcctttc tctcggcctt agcgccattt ttttgggtga gtgttttttg
     2161 gttcctgcgt tgggattccg tgtacaatcc atagacatct gacctcggca cttagcatca
     2221 tcacagcaaa ctaactgtag cctttctctc tttccctgta gaaacctctg cacctgaggc
     2281 caccatgtct agactggaca agagcaaagt cataaacggc gctctggaat tactcaatgg
     2341 agtcggtatc gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc
     2401 taccctgtac tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat
     2461 gctggacagg catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct
     2521 gcggaacaac gccaagtcat tccgctgtgc tctcctctca catcgcgacg gggctaaagt
     2581 gcatctcggc acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt
     2641 cctgtgtcag caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca
     2701 ctttacactg ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga
     2761 gacacctacc accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgaccg
     2821 gcagggagcc gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa
     2881 acagctaaag tgcgaaagcg gcgggccggc cgacgccctt gacgattttg acttagacat
     2941 gctcccagcc gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga
     3001 cgattttgac cttgacatgc tccccgggtc cggcgctact aacttcagcc tgctgaagca
     3061 ggctggtgac gtcgaggaga atcctggtcc catggccatg cctttgtctc aagaagaatc
     3121 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     3181 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     3241 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     3301 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     3361 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     3421 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     3481 ggagggctaa ttcgaaggcc tgtcgtgaag cttggggatc aattctctag agctcgctga
     3541 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3601 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3661 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3721 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3781 gaggcggaaa gaaccagctg ggcctactag ctactcggga ccccttaccg aaacatcgcc
     3841 gcattctgca gaggagtcga gtgtatgtaa acttctgacc cactgggaat gtgatgaaag
     3901 aaataaaagc tgaaatgaat cattctctct actattattc tgatatttca cattcttaaa
     3961 ataaagtggt gatcctaact gacctaagac agggaatttt tactaggatt aaatgtcagg
     4021 aattgtgaaa aagtgagttt aaatgtattt ggctaaggtg tatgtaaact tccgacttca
     4081 actgtatagg gatccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4141 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4201 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4261 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4321 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4381 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4441 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4501 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4561 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4621 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4681 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4741 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4801 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4861 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4921 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4981 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5041 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5101 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5161 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5221 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5281 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5341 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5401 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5461 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5521 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5581 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5641 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5701 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5761 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5821 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5881 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5941 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6001 tttccccgaa aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6061 ataccgcatc aggaaattgt aagcgttaat attttgttaa aatt
//
