LOCUS       sequence-163074-        6858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of myc epitope and murine mRNA Capping Enzyme in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  AUTHORS   Otsuka Y, Kedersha NL, Schoenberg DR
  TITLE     Identification of a cytoplasmic complex that adds a cap onto 
            5'-monophosphate RNA.
  JOURNAL   Mol Cell Biol. 2009 Apr;29(8):2155-67. doi: 10.1128/MCB.01325-08. 
            Epub 2009 Feb 17.
  PUBMED    19223470
REFERENCE   2  (bases 1 to 6858)
  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: 
            "10.1128/MCB.01325-08"; journalName: "Mol Cell Biol"; date: 
            "2009-04"; volume: "29"; issue: "8"; pages: "2155-67"
COMMENT     Sequence Label: pcDNA4/TO-myc-mCE
FEATURES             Location/Qualifiers
     source          1..6858
                     /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"
     primer_bind     complement(2866..2883)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2872..3096
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3142..3570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3229..3248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3439..3460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3579..3599)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3584..3913
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3764..3899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3826..3845
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3961..4008
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4027..4401
                     /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    4531..4652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4568..4587)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4622..4641
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4701..4717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4701..4717)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4714..4736)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4725..4741
                     /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(4749..4755,4756..4773,4774..4779))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4794..4815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4932..4949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5103..5691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5183..5202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5862..6653,6654..6722))
                     /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     6485..6504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6723..6827)
                     /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 atggaggccg cttacaacaa gatcccgccg cggtggctga
     1081 actgtccgcg gcgcggtcag ccggtggcag gaagattctt acctctgaag acgatgctgg
     1141 ggccaagata tgatagtcaa gttgctgaag aaaaccggtt ccatcccagt atgctctcaa
     1201 attatctaaa gagcctgaag gttaaaatga gcttgttggt ggatctgaca aatacttcaa
     1261 ggttctatga cagaaatgac atagaaaaag aaggaatcaa gtatatcaaa cttcagtgta
     1321 aaggacatgg cgaatgcccc accactgaga atactgagac attcattcgc ctgtgtgaga
     1381 ggtttaatga aagaagccca cctgaactta taggtgttca ctgcacccat ggcttcaacc
     1441 gcactggttt ccttatatgt gcctttttgg tggaaaaaat ggattggagt attgaagcag
     1501 cagttgctac atttgcccaa gccagaccac caggaatcta caagggtgat tatttgaagg
     1561 aactttttcg tcgctatgga gatatagaag aagcaccacc accacctgta ttgccagatt
     1621 ggtgttttga agatgaggat gaagaagatg aggatgaaga tggaaaaaag gactcagaac
     1681 cagggtcaag tgcatccttt agtaaaagga gaaaagagcg gttaaaactg ggagctattt
     1741 tcttggaagg cataactgtt aaaggtgtaa ctcaagtaac aactcaaccc aagttaggag
     1801 aggtgcagca gaaatgtcat cagttctgtg gctgggaagg gtctgggttc cctggagcac
     1861 agcctgtctc catggacaag caaaatatta gacttttaga gcagaagcct tataaagtaa
     1921 gctggaaagc agacggcact cgttacatga tgttgattga tggcacaaat gaagttttta
     1981 tgattgacag agataattcc gtttttcatg tttcaaatct ggaatttcca tttcgtaaag
     2041 atcttcgcat gcatttatca aatactcttt tggatgggga aatgatcatt gacaaagtaa
     2101 atggacaggc cgtcccaaga tatttgatct atgacataat taagttcaat gcacaaccag
     2161 ttggagactg tgattttaat attcgtctac agtgtattga acgtgaaatt ataagtccac
     2221 gacatgaaaa aatgaagact ggactcattg acaaaacaca ggaaccattt agtgtcagac
     2281 ctaaacaatt ttttgacatc aatatttcaa gaaagcttct ggaaggaaat tttgccaaag
     2341 aagtcagcca tgaaatggat ggacttattt ttcagcctat tggaaaatac aagcctggtc
     2401 gatgtgatga cattttgaaa tggaaacctc ccagtctaaa ctctgtggat tttcgactta
     2461 agataacaag aatgggagga gaagggttgc ttccacagaa tgttggcctt ctctatgttg
     2521 gaggttatga aagacccttt gcacaaatca aggtgacaaa agaactaaaa cagtatgaca
     2581 acaaaattat agaatgcaaa tttgagaaca atagctgggt cttcatgaga cagaggatag
     2641 acaaaagttt cccaaatgcc tacaacacag ccatggctgt gtgcaatagc atctcgaacc
     2701 ctgtcaccaa ggagatgctg tttgaattca ttgacagatg tgcagcagcc gcccagggac
     2761 agaagcggaa gtatcccctg gaccctgaca cggagctcat gccgccccca ccgcccaaaa
     2821 gactgcatcg gccaacctga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     2881 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2941 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3001 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3061 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3121 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3181 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3241 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3301 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3361 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3421 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3481 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3541 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3601 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3661 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3721 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3781 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3841 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3901 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     3961 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     4021 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     4081 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     4141 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     4201 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     4261 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
     4321 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
     4381 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
     4441 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4501 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4561 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4621 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4681 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4741 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4801 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4861 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4921 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4981 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5041 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5281 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5521 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6001 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6601 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6721 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6841 aaagtgccac ctgacgtc
//
