LOCUS       sequence-429978-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible cpTurboID (V5) for Sleeping Beauty transposon system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7070)
  AUTHORS   Kim JW, Yong AJH, Aisenberg EE, Lobel JH, Wang W, Dawson TM, Dawson 
            VL, Gao R, Jan YN, Bateup HS, Ingolia NT
  TITLE     Molecular recording of calcium signals via calcium-dependent 
            proximity labeling.
  JOURNAL   Nat Chem Biol. 2024 Jul;20(7):894-905. doi: 
            10.1038/s41589-024-01603-7. Epub 2024 Apr 24.
  PUBMED    38658655
REFERENCE   2  (bases 1 to 7070)
  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"; doi: 
            "10.1038/s41589-024-01603-7"; journalName: "Nat Chem Biol"; date: 
            "2024-07"; volume: "20"; issue: "7"; pages: "894-905"
COMMENT     Sequence Label: pNTI822
FEATURES             Location/Qualifiers
     source          1..7070
                     /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(689..697,698..712)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1700..1741
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1780..1901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1791..1810)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1845..1864
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             join(2624..3241,3242..3247,3248..3283,3284..3286,3287..3322,
                     3323..3325,3326..3361,3362..3367)
                     /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             3374..3430
                     /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             3431..4456
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3431..4456
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(4511..4528)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4517..4741
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5067..5084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5238..5826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5318..5337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5997..6788,6789..6857))
                     /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"
     promoter        complement(6858..6962)
                     /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 ccaccatgga tctgtacgac gatgacgata agctcgaggg
      721 gagcgtggca gtcctgcctg tggtcgactc caccaatcag tacctgctgg atcgaatcgg
      781 cgagctgaag agtggggatg cttgcattgc agaatatcag caggcaggga gaggaagcag
      841 agggaggaaa tggttctctc cttttggagc taacctgtac ctgagtatgt tttggcgcct
      901 gaagcgggga ccagcagcaa tcggcctggg cccggtcatc ggaattgtca tggcagaagc
      961 gctgcgaaag ctgggagcag acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca
     1021 ggatagaaag ctggcaggca tcccggtgga gctggccgga ataacaggcg atgctgcaca
     1081 gatcgtcatt ggcgccggga ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa
     1141 tcagggctgg atcacactgc aggaagcagg gattaacctg gacaggaata ctctggccgc
     1201 tacgctgatc cgagagctgc gggcagccct ggaactgttc gagcaggaag gcctggctcc
     1261 atatctgcca cggtgggaga agctggataa cttcatcaat agacccgtga agctgatcat
     1321 tggggacaaa gagattttcg ggattagccg ggggattgat aaacagggag ccctgctgct
     1381 ggaacaggac ggagttatca aaccctggat gggcggagaa atcagtctgc ggtctgccga
     1441 aaagctgcag ggcggtaccg gagggagcaa agacaatact gtgcctctga agctgatcgc
     1501 tctcctggct aatggcgagt tccatagtgg cgaacagctg ggagaaaccc tgggcatgtc
     1561 cagggccgct atcaacaagc acattcagac tctgcgcgac tggggcgtgg acgtgttcac
     1621 cgtgcccgga aagggctact ctctgcccga gcctatcccg ctgctgaacg ctaaacagat
     1681 tctgggacag ctggacggcg gcaagcccat ccccaacccc ctgctgggcc tggacagcac
     1741 ctgaggcctg tcaggccaag cttccatcga tagacatgat aagatacatt gatgagtttg
     1801 gacaaaccac aacaagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1861 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1921 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     1981 acaaatgtgg tacctaggcg agaccctgtc tcacaaaata aagtaagccc ggactgagtg
     2041 cggaaaggcg ggcctggcgg gtctggtctc cccatgcggg ccaccagagg ccctgcagcc
     2101 ttcagtcgct tgaaggggta atggcgcttc cactcacaaa catggcggac agagcgtgtg
     2161 aacgagatga acagcccctc aaaaatatgg ccgccgaggc tggacggccg tgccccagca
     2221 gcaccgcctc cgcgccccac gtgatctctc gccgggcaca gcgctgaccg cggaggtcca
     2281 accggaagaa tgtccggatt ggacattcgg aagagggccc gccttccctg gggaatctct
     2341 gcgcacgcgc agaacgcttc gaccaatgaa aacacaggaa gccgtccgcg caaccgcgtt
     2401 gcgtcacttc tgccgcccct gtttcaaggt atatagccgt agacggaact tcgcctttct
     2461 ctcggcctta gcgccatttt tttgggtgag tgttttttgg ttcctgcgtt gggattccgt
     2521 gtacaatcca tagacatctg acctcggcac ttagcatcat cacagcaaac taactgtagc
     2581 ctttctctct ttccctgtag aaacctctgc acctgaggcc accatgtcta gactggacaa
     2641 gagcaaagtc ataaacggcg ctctggaatt actcaatgga gtcggtatcg aaggcctgac
     2701 gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa
     2761 gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca
     2821 cttctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcatt
     2881 ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac
     2941 agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc
     3001 cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt
     3061 ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat
     3121 gcccccactt ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt
     3181 ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg
     3241 cgggccggcc gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga
     3301 cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct
     3361 ccccgggtcc ggcgctacta acttcagcct gctgaagcag gctggtgacg tcgaggagaa
     3421 tcctggtccc atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     3481 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     3541 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     3601 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     3661 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     3721 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     3781 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     3841 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     3901 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     3961 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     4021 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     4081 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     4141 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     4201 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     4261 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     4321 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     4381 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     4441 gagggcaaag gaatagttcg aaggcctgtc gtgaagcttg gggatcaatt ctctagagct
     4501 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4561 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4621 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4681 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4741 gcttctgagg cggaaagaac cagctgggcc tactagctac tcgggacccc ttaccgaaac
     4801 atcgccgcat tctgcagagg agtcgagtgt atgtaaactt ctgacccact gggaatgtga
     4861 tgaaagaaat aaaagctgaa atgaatcatt ctctctacta ttattctgat atttcacatt
     4921 cttaaaataa agtggtgatc ctaactgacc taagacaggg aatttttact aggattaaat
     4981 gtcaggaatt gtgaaaaagt gagtttaaat gtatttggct aaggtgtatg taaacttccg
     5041 acttcaactg tatagggatc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5101 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5161 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5221 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5281 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5341 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5401 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5461 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5521 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5581 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5641 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5701 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5761 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5821 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5881 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5941 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6001 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6061 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6121 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6181 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6241 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6301 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6361 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6421 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6481 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6541 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6601 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6661 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6721 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6781 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6841 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6901 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6961 cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag
     7021 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
//
