LOCUS       sequence-204869-        6261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA - TET DBD - MTAD. Expresses TET DNA binding domain fused to 
            MTAD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6261)
  AUTHORS   Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS, 
            Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP
  TITLE     Biosynthesis of Orthogonal Molecules Using Ferredoxin and 
            Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded 
            PhyB Optogenetics.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 
            10.1021/acssynbio.7b00413. Epub 2018 Jan 24.
  PUBMED    29301067
REFERENCE   2  (bases 1 to 6261)
  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: 
            "10.1021/acssynbio.7b00413"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "706-717"
COMMENT     Sequence Label: pPKm-293
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     655..674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             696..716
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             720..1340
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(793..812)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             1527..1538
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1558..1578
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1606..1624)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1607..1624)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1644..1661)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1650..1874
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1920..2348
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2007..2026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2217..2238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2357..2377)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2362..2691
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2542..2677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2604..2623
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2758..3552
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2812..2831)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3422..3441
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3726..3847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3763..3782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3817..3836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3896..3912)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3896..3912)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3909..3931)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3920..3936
                     /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(3944..3950,3951..3968,3969..3974))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3989..4010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4127..4144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4298..4886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4378..4397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5057..5848,5849..5917))
                     /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     5680..5699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5918..6022)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6097..6116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagcttggta ccatgcctaa gaaaaagaga aaggtgggca
      721 tgtcaagatt agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg
      781 gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat
      841 tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag
      901 ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta
      961 ataacgctaa aagttttaga tgtgctttac taagtcatcg ggatggagca aaagtacatt
     1021 taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat
     1081 gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg gggcatttta
     1141 ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac
     1201 ctactactga tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag
     1261 gtgcagagcc agccttctta ttcggccttg aattgatcat ctgcggatta gaaaaacaac
     1321 ttaaatgtga aagtgggtcc ggatccatgg gatctcctag gtttgatact gacctgttcg
     1381 ataccgactt gtttgatact gacctttttg atactgacct gttcgatacc gacttgtttg
     1441 atactgacct ttttgatact gacctgttcg ataccgactt gtttgatact gacctttttg
     1501 atactgacct gttcgatacc gacttgatcg agggaagggg tggcgaatta attcatgcct
     1561 aagaaaaaga gaaaggtggg cggaggcgct agtagagggc cctattctat agtgtcacct
     1621 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1681 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1741 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1801 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     1861 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     1921 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1981 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2041 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2101 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2161 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2221 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2281 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2341 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2401 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2461 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2521 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2581 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2641 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     2701 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     2761 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     2821 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     2881 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     2941 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3001 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3061 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3121 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3181 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3241 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3301 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3361 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3421 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3481 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3541 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3601 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3661 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3721 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3781 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3841 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3901 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3961 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4021 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4081 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4141 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4201 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4261 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4321 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4381 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4441 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4501 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4561 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4621 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4681 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4741 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4801 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4861 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4921 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4981 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5041 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5101 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5161 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5221 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5281 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5341 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5401 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5461 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5521 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5581 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5641 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5701 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5761 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5821 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5881 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5941 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6001 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6061 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     6121 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     6181 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     6241 taggcgtttt gcgctgcttc g
//
