LOCUS       sequence-326691-        8101 bp    DNA     circular SYN 15-APR-2025
DEFINITION  rtTA under the control of PGK1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8101)
  AUTHORS   Gligorovski V, Sadeghi A, Rahi SJ
  TITLE     Multidimensional characterization of inducible promoters and a 
            highly light-sensitive LOV-transcription factor.
  JOURNAL   Nat Commun. 2023 Jun 27;14(1):3810. doi: 10.1038/s41467-023-38959-8.
  PUBMED    37369667
REFERENCE   2  (bases 1 to 8101)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-06-27"; volume: "14(1):3810. doi"; pages: " 
            10.1038/s41467-023-38959-8"
COMMENT     Sequence Label: pVG48
FEATURES             Location/Qualifiers
     source          1..8101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1384..2004,2005..2391)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MPRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(1457..1476)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2405..2652
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2421..2439)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(4119..4306)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        complement(4632..4650)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4671..4687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4671..4687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4684..4706)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4695..4711
                     /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)."
     promoter        complement(join(4719..4725,4726..4743,4744..4749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4764..4785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4902..4919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5073..5661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5153..5172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5832..6623,6624..6692))
                     /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     6455..6474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6693..6797)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6881..6899
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6937..6959)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7059..7078
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7434..7889)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7571..7592)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7783..7802
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8015..8037
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8029..8046
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8030..8046
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8053..8072
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8053..8071
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggccggccgc tgccaccgtt gtttccaccg aagaaaccac cgttgccgta accaccacga
       61 cggttgttgc taaagaagct gccaccgcca cggccaccgt tgtagccgcc gttgttgtta
      121 ttgtagttgc tactgttatt tctggcactt cttggttttc ctcttaagtg aggaggaaca
      181 taaccattct cgttgttgtc gttgatgctt aaattttgca cttgttcgct cagttcagcc
      241 ataatatgaa atgcttttct tgttgttctt acggaatacc acttgccacc tatcaccaca
      301 actaactttt tcccgttcct ccatctcttt tatatttttt ttctggagtt caagagaaaa
      361 aaaaagaaaa agcaaaaaga aaaaaggaaa gcgcgcctcg ttcagaatga cacgtataga
      421 atgatgcatt accttgtcat cttcagtatc atactgttcg tatacatact tactgacatt
      481 cataggtata catatataca catgtatata tatcgtatgc tgcagcttta aataatcggt
      541 gtcaagatct gagctccacc gcggtggcgg ccgcttattt actcaggatc ccgcatgttg
      601 caggtgggag ttccagctcc cgggaaacag ctgcaagaat tactcgtgag taaggaaaga
      661 gtgaggaact atcgcatacc tgcatttaaa gatgccgatt tgggcgcgaa tcctttattt
      721 tggcttcacc ctcatactat tatcagggcc agaaaaagga agtgtttccc tccttcttga
      781 attgatgtta ccctcataaa gcacgtggcc tcttatcgag aaagaaatta ccgtcgctcg
      841 tgatttgttt gcaaaaagaa caaaactgaa aaaacccaga cacgctcgac ttcctgtctt
      901 cctattgatt gcagcttcca atttcgtcac acaacaaggt cctagcgacg gctcacaggt
      961 tttgtaacaa gcaatcgaag gttctggaat ggcgggaaag ggtttagtac cacatgctat
     1021 gatgcccact gtgatctcca gagcaaagtt cgttcgatcg tactgttact ctctctcttt
     1081 caaacagaat tgtccgaatc gtgtgacaac aacagcctgt tctcacacac tcttttcttc
     1141 taaccaaggg ggtggtttag tttagtagaa cctcgtgaaa cttacattta catatatata
     1201 aacttgcata aattggtcaa tgcaagaaat acatatttgg tcttttctaa ttcgtagttt
     1261 ttcaagttct tagatgcttt ctttttctct tttttacaga tcatcaagga agtaattatc
     1321 tactttttac aacaaatata aaacaccgcg gtggcggccg ctaaaatcat ggctagcgga
     1381 tccatgccta gattagataa aagtaaagtg attaacagcg cattagagct gcttaatggg
     1441 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
     1501 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttacc cattgagatg
     1561 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
     1621 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
     1681 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
     1741 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcac
     1801 tttactttag gttgcgtatt ggaagaacaa gagcatcaag tcgctaaaga agaaagggaa
     1861 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcgc
     1921 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
     1981 caacttaaat gtgaaagtgg gtccgcgtac agccgcgcgc gtacgaaaaa caattacggg
     2041 tctaccatcg agggcctgct cgatctcccg gacgacgacg cccccgaaga ggcggggctg
     2101 gcggctccgc gcctgtcctt tctccccgcg ggacacacgc gcagactgtc gacggccccc
     2161 ccgaccgatg tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg
     2221 catgccgacg cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt
     2281 ccgggattta ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag
     2341 tttgagcaga tgtttaccga tgcccttgga attgacgagt acggtgggta ggctcgagta
     2401 atgatcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
     2461 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
     2521 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
     2581 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
     2641 gctttaattt gcggccttat cgccggcgaa taaattaagg cgcgccaaca tatccggagg
     2701 tacccaaagc cgaatccacc acggtcaccg ttgtttctga agaaaccacc acggccgcca
     2761 ccgttgttga aaccaccacg aggagcagaa tcgcctcttc tgccaccacc gttacggatg
     2821 tgaggtggaa cgtaaccacc acgtctttgt tgcccttcag gcatggaaac ttgctctgct
     2881 aactcagtca tattggatat atgtattact ttctttattt tttgtagttt agtcttctat
     2941 ttcttttata gtttggtatc tttttgcaaa aaatttacct ctatatcgtg ttcgtgcgta
     3001 tttcttcctc tataatatga aaagaaaggc agtagtcttg gttttctatt attccaaata
     3061 ttacaagaat acccagaacc acaagaagca gaagaaacta actgctttat atacttaatt
     3121 tcccgccaca agctcatcgc ccagaagttt ttttcagttt ttcacctgac aatctggcag
     3181 ctcgctgaaa aattttcacc tggggtcgcc tgctacgtag agtgatcacg tttatgaata
     3241 gcacattata tagtggccca gacggtctga tggtagttca tgtagttaag aataaatact
     3301 ctgtttgtgt gatttctatg tgtacgttat ctatataaat attgttgatg gtggtagctg
     3361 tgggctgtgt tctatgctag gacaccctta gtggatggaa cgtcatcgcg acccgtaggt
     3421 gttcttgcct ctctgatggc gatggcgagc gccttgaaag ctgattcgga gcggtggtgg
     3481 tcgttggtgc cccgcagaca gtccacatgc atggtgatac gcgcctcagt ggcgaaactc
     3541 tccaagaagt gcgggatcat ctctgtggat agctggccta tgcgttctcg cttcaggccc
     3601 agctccacca cggcgaaagg cctattggag aagtcaacca cagcgcggct cagcgcctcg
     3661 tccagtggtg caaacccatg cccgaacctc ttgataccac gcacggagcc aagcgcctcc
     3721 ttgaacgctt ggcccagcgc aatcccacag tcctcaacag tgtggtggtc atcgatgtgc
     3781 aagtccccta tacactccag gatcagagac cacccgccat gcttcgccag cgcagtcaac
     3841 atgtggtcca aaaacccaac tccagtgtgc acatggatgg tctgcccatc accactagca
     3901 tattccttgc ccagtatgct ctcccgtaca gaaacagacc caccgtcaag atccagcgcc
     3961 agctgtatat tcgtctcctg cgtaaccctc ttaacaaacg ccatcctttg ctcgatgctt
     4021 ctctttgctt caattatctc aactttaagt aaacctgaag taaactagaa ctagattaaa
     4081 tcagaaggca tccgcagctt tatatacata tagagcctct ttgccttcgt ttatcttgcc
     4141 tgctcatttt ttagtatatt cttcgaagaa atcacattac tttatataat gtataattca
     4201 ttatgtgata atgccaatcg ctaagaaaaa aaaagagtca tccgctaggt ggaaaaaaaa
     4261 aaatgaaaat cattaccgag gcataaaaaa atatagagtg tactagagga ggccaagagt
     4321 aatagaaaaa gaaaattgcg ggaaaggact gtgttatgac ttccctgact aatgccgtgt
     4381 tcaaacgata cctggcagtg actcctagcg ctcaccaagc tcttaaaacg agaattaaga
     4441 aaaagtcgtc atctttcgat aagtttttcc cacagcaaag caatagtaga aaaaaacaat
     4501 gggaaacgtt gaatgaagac aaagcgtcgt ggtttaaaag gaaatacgct cacgtacatg
     4561 ctagggaaca ggaccgtgca gcggccagcg tggccgttta aacatttaaa tggccggccc
     4621 agcttttgtt ccctttagtg agggttaatt ccgagcttgg cgtaatcatg gtcatagctg
     4681 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4741 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4801 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4861 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5881 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5941 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6001 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6061 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6121 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6181 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6241 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6301 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6361 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6421 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6481 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6541 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6601 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6661 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6721 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6781 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcct cggcgcgtga
     6841 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6901 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6961 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     7021 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     7081 gagagtgcac cataccacct tttcaattca tcattttttt tttattcttt tttttgattt
     7141 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
     7201 gagcacagac ttagattggt atatatacgc atattgggtc agcagtacag aaccctcgac
     7261 gaagttgtaa gtggtggatc agatatcatc attgttggca gaggactttt cgccaagggt
     7321 agagatccta atactaaact cacaaattag agcttcaatt taattatatc agttattacc
     7381 ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     7441 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     7501 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     7561 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7621 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7681 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7741 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7801 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7861 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     7921 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7981 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     8041 gccagtgaat tgtaatacga ctcactatag ggggccagcg tggccgttta aacatttaaa
     8101 t
//
