LOCUS       sequence-217051-        7195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For tetracycline-inducible expression of bio-myc-NES-mCE Delta-NLS 
            (""bio-myc-cCE"") in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7195)
  AUTHORS   Trotman JB, Agana BA, Giltmier AJ, Wysocki VH, Schoenberg DR
  TITLE     RNA-binding proteins and heat-shock protein 90 are constituents of 
            the cytoplasmic capping enzyme interactome.
  JOURNAL   J Biol Chem. 2018 Oct 26;293(43):16596-16607. doi: 
            10.1074/jbc.RA118.004973. Epub 2018 Aug 30.
  PUBMED    30166341
REFERENCE   2  (bases 1 to 7195)
  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: 
            "10.1074/jbc.RA118.004973"; journalName: "J Biol Chem"; date: 
            "2018-10-26- 26"; volume: "293"; issue: "43"; pages: "16596-16607"
COMMENT     Sequence Label: pcDNA4/TO-bio-myc-cCE
FEATURES             Location/Qualifiers
     source          1..7195
                     /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"
     protein_bind    612..630
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    633..651
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     661..685
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1121..1147
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2943..2966
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2995..3012)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3001..3225
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3271..3699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3358..3377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3568..3589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3708..3728)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3713..4042
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3893..4028
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3955..3974
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4090..4137
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4156..4530
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4660..4781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4697..4716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4751..4770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4830..4846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4830..4846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4843..4865)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4854..4870
                     /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(4878..4884,4885..4902,4903..4908))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4923..4944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5061..5078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5232..5820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5312..5331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5991..6782,6783..6851))
                     /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     6614..6633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6852..6956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7031..7050)
                     /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 ctccctatca gtgatagaga tctccctatc agtgatagag atcgtcgacg
      661 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
      721 tagaagacac cgggaccgat ccagcctccg gactctagcg tttaaactta agcttggtac
      781 catggagtta aaaaccgccg cactagctca acatgacgag gctgctggaa aggccggtga
      841 aggtgaaatc cctgcccctc ttgctggtac cgtttctaag atactggtaa aagaaggtga
      901 cactgttaaa gctggtcaaa cagttctggt gctggaggct atgaaaatgg agacagaaat
      961 taacgctcct actgacggaa aagttgaaaa ggtgttagtt aaggaaagag atgctgttca
     1021 aggtggtcaa ggtctaatca agatcggcgt tggcaccatg gcatcaatgc agaagctgat
     1081 ctcagaggag gacctgctta tggccatgga ggcccttcag ctaccaccgc ttgagagact
     1141 tactcttgat atggccatgg aggccgctta caacaagatc ccgccgcggt ggctgaactg
     1201 tccgcggcgc ggtcagccgg tggcaggaag attcttacct ctgaagacga tgctggggcc
     1261 aagatatgat agtcaagttg ctgaagaaaa ccggttccat cccagtatgc tctcaaatta
     1321 tctaaagagc ctgaaggtta aaatgagctt gttggtggat ctgacaaata cttcaaggtt
     1381 ctatgacaga aatgacatag aaaaagaagg aatcaagtat atcaaacttc agtgtaaagg
     1441 acatggcgaa tgccccacca ctgagaatac tgagacattc attcgcctgt gtgagaggtt
     1501 taatgaaaga agcccacctg aacttatagg tgttcactgc acccatggct tcaaccgcac
     1561 tggtttcctt atatgtgcct ttttggtgga aaaaatggat tggagtattg aagcagcagt
     1621 tgctacattt gcccaagcca gaccaccagg aatctacaag ggtgattatt tgaaggaact
     1681 ttttcgtcgc tatggagata tagaagaagc accaccacca cctgtattgc cagattggtg
     1741 ttttgaagat gaggatgaag aagatgagga tgaagatgga aaaaaggact cagaaccagg
     1801 gtcaagtgca tcctttagta aaaggagaaa agagcggtta aaactgggag ctattttctt
     1861 ggaaggcata actgttaaag gtgtaactca agtaacaact caacccaagt taggagaggt
     1921 gcagcagaaa tgtcatcagt tctgtggctg ggaagggtct gggttccctg gagcacagcc
     1981 tgtctccatg gacaagcaaa atattagact tttagagcag aagccttata aagtaagctg
     2041 gaaagcagac ggcactcgtt acatgatgtt gattgatggc acaaatgaag tttttatgat
     2101 tgacagagat aattccgttt ttcatgtttc aaatctggaa tttccatttc gtaaagatct
     2161 tcgcatgcat ttatcaaata ctcttttgga tggggaaatg atcattgaca aagtaaatgg
     2221 acaggccgtc ccaagatatt tgatctatga cataattaag ttcaatgcac aaccagttgg
     2281 agactgtgat tttaatattc gtctacagtg tattgaacgt gaaattataa gtccacgaca
     2341 tgaaaaaatg aagactggac tcattgacaa aacacaggaa ccatttagtg tcagacctaa
     2401 acaatttttt gacatcaata tttcaagaaa gcttctggaa ggaaattttg ccaaagaagt
     2461 cagccatgaa atggatggac ttatttttca gcctattgga aaatacaagc ctggtcgatg
     2521 tgatgacatt ttgaaatgga aacctcccag tctaaactct gtggattttc gacttaagat
     2581 aacaagaatg ggaggagaag ggttgcttcc acagaatgtt ggccttctct atgttggagg
     2641 ttatgaaaga ccctttgcac aaatcaaggt gacaaaagaa ctaaaacagt atgacaacaa
     2701 aattatagaa tgcaaatttg agaacaatag ctgggtcttc atgagacaga ggatagacaa
     2761 aagtttccca aatgcctaca acacagccat ggctgtgtgc aatagcatct cgaaccctgt
     2821 caccaaggag atgctgtttg aattcattga cagatgtgca gcagccgccc agggacagcc
     2881 cctggaccct gacacggagc tcatgccgcc cccaccgccc aaaagactgc atcggccaac
     2941 ctgattacaa ggatgacgac gataagtagg ggcccgttta aacccgctga tcagcctcga
     3001 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3061 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3121 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3181 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3241 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3301 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3361 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3421 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3481 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3541 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3601 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3661 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3721 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3781 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3841 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3901 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3961 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4021 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4081 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     4141 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     4201 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     4261 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     4321 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     4381 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     4441 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     4501 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     4561 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4621 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4681 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4741 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4801 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4861 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4921 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4981 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5041 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5101 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5401 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5641 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5941 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6001 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6061 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6961 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     7021 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     7081 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     7141 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcg
//
