LOCUS       sequence-163076-        7627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cytoplasmic restricted form of N-terminal myc tagged and 
            C-terminal biotin tagged mRNA capping enzyme, inactive form.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7627)
  AUTHORS   Mukherjee C, Bakthavachalu B, Schoenberg DR
  TITLE     The cytoplasmic capping complex assembles on adapter protein nck1 
            bound to the proline-rich C-terminus of Mammalian capping enzyme.
  JOURNAL   PLoS Biol. 2014 Aug 19;12(8):e1001933. doi: 
            10.1371/journal.pbio.1001933. eCollection 2014 Aug.
  PUBMED    25137142
REFERENCE   2  (bases 1 to 7627)
  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: "PLoS 
            Biol."; date: "2014-08-19"; volume: "12(8):e1001933. doi"; pages: " 
            10.1371/journal.pbio.1001933. eCollection 2014 Aug"
COMMENT     Sequence Label: pcDNA3/ TO myc-cCE-bio (cCE-bio)
FEATURES             Location/Qualifiers
     source          1..7627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1056..1082
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2877..2897
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(3180..3198)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3181..3198)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3218..3235)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3224..3448
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3494..3922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3581..3600)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3791..3812
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3931..3951)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3936..4265
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4116..4251
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4178..4197
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4332..5126
                     /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(4386..4405)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4996..5015
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5300..5421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5337..5356)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5391..5410
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5470..5486)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5470..5486)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5483..5505)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5494..5510
                     /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(5518..5524,5525..5542,5543..5548))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5563..5584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5701..5718)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5872..6460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5952..5971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6631..7422,7423..7491))
                     /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     7254..7273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7492..7596)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca ccatggcatc aatgcagaag ctgatctcag
     1021 aggaggacct gcttatggcc atggaggccc ttcagctacc accgcttgag agacttactc
     1081 ttgatatggc catggaggcc gcttacaaca agatcccgcc gcggtggctg aactgtccgc
     1141 ggcgcggtca gccggtggca ggaagattct tacctctgaa gacgatgctg gggccaagat
     1201 atgatagtca agttgctgaa gaaaaccggt tccatcccag tatgctctca aattatctaa
     1261 agagcctgaa ggttaaaatg agcttgttgg tggatctgac aaatacttca aggttctatg
     1321 acagaaatga catagaaaaa gaaggaatca agtatatcaa acttcagtgt aaaggacatg
     1381 gcgaatgccc caccactgag aatactgaga cattcattcg cctgtgtgag aggtttaatg
     1441 aaagaagccc acctgaactt ataggtgttc actgcaccca tggcttcaac cgcactggtt
     1501 tccttatatg tgcctttttg gtggaaaaaa tggattggag tattgaagca gcagttgcta
     1561 catttgccca agccagacca ccaggaatct acaagggtga ttatttgaag gaactttttc
     1621 gtcgctatgg agatatagaa gaagcaccac caccacctgt attgccagat tggtgttttg
     1681 aagatgagga tgaagaagat gaggatgaag atggaaaaaa ggactcagaa ccagggtcaa
     1741 gtgcatcctt tagtaaaagg agaaaagagc ggttaaaact gggagctatt ttcttggaag
     1801 gcataactgt taaaggtgta actcaagtaa caactcaacc caagttagga gaggtgcagc
     1861 agaaatgtca tcagttctgt ggctgggaag ggtctgggtt ccctggagca cagcctgtct
     1921 ccatggacaa gcaaaatatt agacttttag agcagaagcc ttataaagta agctggaaag
     1981 cagacggcac tcgttacatg atgttgattg atggcacaaa tgaagttttt atgattgaca
     2041 gagataattc cgtttttcat gtttcaaatc tggaatttcc atttcgtaaa gatcttcgca
     2101 tgcatttatc aaatactctt ttggatgggg aaatgatcat tgacaaagta aatggacagg
     2161 ccgtcccaag atatttgatc tatgacataa ttaagttcaa tgcacaacca gttggagact
     2221 gtgattttaa tattcgtcta cagtgtattg aacgtgaaat tataagtcca cgacatgaaa
     2281 aaatgaagac tggactcatt gacaaaacac aggaaccatt tagtgtcaga cctaaacaat
     2341 tttttgacat caatatttca agaaagcttc tggaaggaaa ttttgccaaa gaagtcagcc
     2401 atgaaatgga tggacttatt tttcagccta ttggaaaata caagcctggt cgatgtgatg
     2461 acattttgaa atggaaacct cccagtctaa actctgtgga ttttcgactt aagataacaa
     2521 gaatgggagg agaagggttg cttccacaga atgttggcct tctctatgtt ggaggttatg
     2581 aaagaccctt tgcacaaatc aaggtgacaa aagaactaaa acagtatgac aacaaaatta
     2641 tagaatgcaa atttgagaac aatagctggg tcttcatgag acagaggata gacaaaagtt
     2701 tcccaaatgc ctacaacaca gccatggctg tgtgcaatag catctcgaac cctgtcacca
     2761 aggagatgct gtttgaattc attgacagat gtgcagcagc cgcccaggga cagcccctgg
     2821 accctgacac ggagctcatg ccgcccccac cgcccaaaag actgcatcgg ccaaccgaga
     2881 atctgtactt tcaatcagag ttaaaaaccg ccgcactagc tcaacatgac gaggctgctg
     2941 gaaaggccgg tgaaggtgaa atccctgccc ctcttgctgg taccgtttct aagatactgg
     3001 taaaagaagg tgacactgtt aaagctggtc aaacagttct ggtgctggag gctatgaaaa
     3061 tggagacaga aattaacgct cctactgacg gaaaagttga aaaggtgtta gttaaggaaa
     3121 gagatgctgt tcaaggtggt caaggtctaa tcaagatcgg cgtttaatga gggccctatt
     3181 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     3241 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3301 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3361 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3421 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3481 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3541 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3601 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3661 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3721 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3781 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3841 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3901 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3961 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4021 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4081 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4141 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4201 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4261 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4321 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4381 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4441 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4501 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4561 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4621 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4681 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4741 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4801 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4861 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4921 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4981 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5041 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5101 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5161 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5221 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5281 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5341 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5401 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5461 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5521 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5581 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5641 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5701 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5761 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5821 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5881 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5941 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6001 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6061 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6121 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6181 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6241 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6301 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6361 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6541 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6601 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6661 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6721 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6781 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6841 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6901 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6961 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7021 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7081 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7141 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7201 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7261 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7321 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7381 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7441 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7501 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7561 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7621 tgacgtc
//
