LOCUS       sequence-163075-        5784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-GFP-Flag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5784)
  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 5784)
  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- GFP-Flag
FEATURES             Location/Qualifiers
     source          1..5784
                     /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             993..1022
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1068..1096)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1691..1711
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1740..1763
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1792..1809)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1798..2022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2068..2496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2155..2174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2365..2386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2505..2525)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2510..2839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2690..2825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2752..2771
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2887..2934
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2953..3327
                     /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    3457..3578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3494..3513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3548..3567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3627..3643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3627..3643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3640..3662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3651..3667
                     /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(3675..3681,3682..3699,3700..3705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3720..3741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3858..3875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4029..4617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4109..4128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4788..5579,5580..5648))
                     /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     5411..5430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5649..5753)
                     /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 tggaacaaaa gctgatttca gaagaagatc
     1021 taaagctgga attcatggct agcaaaggag aagaactttt cactggagtt gtcccaattc
     1081 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     1141 gtgatgctac atacggaaag cttaccctta aatttatttg cactactgga aaactacctg
     1201 ttccatggcc aacacttgtc actactttct cttatggtgt tcaatgcttt tcccgttatc
     1261 cggatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg
     1321 aacgcactat atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg
     1381 aaggtgatac ccttgttaat cgtatcgagt taaaaggtat tgattttaaa gaagatggaa
     1441 acattctcgg acacaaactc gagtacaact ataactcaca caatgtatac atcacggcag
     1501 acaaacaaaa gaatggaatc aaagctaact tcaaaattcg tcacaacatt gaagatggat
     1561 ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     1621 taccagacaa ccattacctg tcgacacaat ctgccctttc gaaagatccc aacgaaaagc
     1681 gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg
     1741 attacaagga tgacgacgat aagtaggggc ccgtttaaac ccgctgatca gcctcgactg
     1801 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1861 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1921 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1981 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2041 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2101 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2161 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2221 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2281 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2341 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2401 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2461 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2521 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2581 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2641 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2701 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2761 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     2821 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     2881 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     2941 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     3001 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     3061 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     3121 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     3181 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     3241 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     3301 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     3361 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3421 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3481 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3541 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3601 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3661 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3721 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3781 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3841 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3901 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3961 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4021 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4081 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4141 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4201 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4261 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4321 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4381 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4441 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4501 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4561 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4621 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4681 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4741 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4801 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4861 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4921 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4981 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5041 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5101 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5161 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5221 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5281 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5341 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5401 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5461 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5521 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5581 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5641 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5761 ccccgaaaag tgccacctga cgtc
//
