LOCUS       sequence-157156-        7473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mRNA capping enzyme containing N terminal biotin and myc 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7473)
  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 7473)
  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-biotin-myc- mCE
FEATURES             Location/Qualifiers
     source          1..7473
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3025..3043)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3026..3043)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3063..3080)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3069..3293
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3339..3767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3426..3445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3636..3657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3776..3796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3781..4111
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3962..4097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4024..4043
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4178..4972
                     /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(4232..4251)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4842..4861
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5146..5267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5183..5202)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5237..5256
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5316..5332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5316..5332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5329..5351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5340..5356
                     /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(5364..5370,5371..5388,5389..5394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5409..5430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5547..5564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5718..6306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5798..5817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6477..7268,7269..7337))
                     /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     7100..7119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7338..7442)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggagtt
      901 aaaaaccgcc gcactagctc aacatgacga ggctgctgga aaggccggtg aaggtgaaat
      961 ccctgcccct cttgctggta ccgtttctaa gatactggta aaagaaggtg acactgttaa
     1021 agctggtcaa acagttctgg tgctggaggc tatgaaaatg gagacagaaa ttaacgctcc
     1081 tactgacgga aaagttgaaa aggtgttagt taaggaaaga gatgctgttc aaggtggtca
     1141 aggtctaatc aagatcggcg tttcaccatg gcatcaatgc agaagctgat ctcagaggag
     1201 gacctgctta tggccatgga ggccgcttac aacaagatcc cgccgcggtg gctgaactgt
     1261 ccgcggcgcg gtcagccggt ggcaggaaga ttcttacctc tgaagacgat gctggggcca
     1321 agatatgata gtcaagttgc tgaagaaaac cggttccatc ccagtatgct ctcaaattat
     1381 ctaaagagcc tgaaggttaa aatgagcttg ttggtggatc tgacaaatac ttcaaggttc
     1441 tatgacagaa atgacataga aaaagaagga atcaagtata tcaaacttca gtgtaaagga
     1501 catggcgaat gccccaccac tgagaatact gagacattca ttcgcctgtg tgagaggttt
     1561 aatgaaagaa gcccacctga acttataggt gttcactgca cccatggctt caaccgtact
     1621 ggtttcctta tatgtgcctt tttggtggaa aaaatggatt ggagtattga agcagcagtt
     1681 gctacatttg cccaagccag accaccagga atctacaagg gtgattattt gaaggaactt
     1741 tttcgtcgct atggagatat agaagaagca ccaccaccac ctgtattgcc agattggtgt
     1801 tttgaagatg aggatgaaga agatgaggat gaagatggaa aaaaggactc agaaccaggg
     1861 tcaagtgcat cctttagtaa aaggagaaaa gagcggttaa aactgggagc tattttcttg
     1921 gaaggcataa ctgttaaagg tgtaactcaa gtaacaactc aacccaagtt aggagaggtg
     1981 cagcagaaat gtcatcagtt ctgtggctgg gaagggtctg ggttccctgg agcacagcct
     2041 gtctccatgg acaagcaaaa tattagactt ttagagcaga agccttataa agtaagctgg
     2101 aaagcagacg gcactcgtta catgatgttg attgatggca caaatgaagt ttttatgatt
     2161 gacagagata attccgtttt tcatgtttca aatctggaat ttccatttcg taaagatctt
     2221 cgcatgcatt tatcaaatac tcttttggat ggggaaatga tcattgacaa agtaaatgga
     2281 caggccgtcc caagatattt gatctatgac ataattaagt tcaatgcaca accagttgga
     2341 gactgtgatt ttaatattcg tctacagtgt attgaacgtg aaattataag tccacgacat
     2401 gaaaaaatga agactggact cattgacaaa acacaggaac catttagtgt cagacctaaa
     2461 caattttttg acatcaatat ttcaagaaag cttctggaag gaaattttgc caaagaagtc
     2521 agccatgaaa tggatggact tatttttcag cctattggaa aatacaagcc tggtcgatgt
     2581 gatgacattt tgaaatggaa acctcccagt ctaaactctg tggattttcg acttaagata
     2641 acaagaatgg gaggagaagg gttgcttcca cagaatgttg gccttctcta tgttggaggt
     2701 tatgaaagac cctttgcaca aatcaaggtg acaaaagaac taaaacagta tgacaacaaa
     2761 attatagaat gcaaatttga gaacaatagc tgggtcttca tgagacagag gatagacaaa
     2821 agtttcccaa atgcctacaa cacagccatg gctgtgtgca atagcatctc gaaccctgtc
     2881 accaaggaga tgctgtttga attcattgac agatgtgcag cagccgccca gggacagaag
     2941 cggaagtatc ccctggaccc tgacacggag ctcatgccgc ccccaccgcc caaaagactg
     3001 catcggccaa cctgagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     3061 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3121 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3181 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3241 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3301 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3361 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3421 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3481 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3541 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3601 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3661 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     3721 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3781 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag
     3841 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3901 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3961 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4021 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4081 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4141 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4201 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4261 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4321 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4381 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4441 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4501 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4561 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4621 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4681 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4741 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4801 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4861 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4921 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4981 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5041 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5101 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5161 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5221 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5281 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5341 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5401 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5461 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5521 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5581 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5641 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5701 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5761 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5821 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5881 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     5941 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6001 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6061 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6121 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6181 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6241 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6301 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6361 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6421 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6481 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6541 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6601 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6661 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6721 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6781 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6841 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6901 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6961 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7021 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7081 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7141 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7201 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7261 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7321 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7381 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7441 cgcacatttc cccgaaaagt gccacctgac gtc
//
