LOCUS       sequence-176338-        8415 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mammalian expression of EDC4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8415)
  AUTHORS   D'Lima NG, Ma J, Winkler L, Chu Q, Loh KH, Corpuz EO, Budnik BA, 
            Lykke-Andersen J, Saghatelian A, Slavoff SA
  TITLE     A human microprotein that interacts with the mRNA decapping complex.
  JOURNAL   Nat Chem Biol. 2016 Dec 5. doi: 10.1038/nchembio.2249.
  PUBMED    27918561
REFERENCE   2  (bases 1 to 8415)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2016 Dec 5. doi: 10.1038/nchembio.2249."
COMMENT     Sequence Label: myc EDC4 pCMVSport6
FEATURES             Location/Qualifiers
     source          1..8415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        204..507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        508..711
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     662..682
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     708..732
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     837..853
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     837..853
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        864..882
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     864..881
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    883..907
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1080..1109
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(5736..5760)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5761..5780)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5762..5780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5791..5808)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5791..5807)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5800..5822)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    6212..6346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6249..6268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6303..6322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6416..6433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6587..7175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6667..6686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(7253..7286)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(7388..8179,8180..8248))
                     /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     8011..8030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctgccgagca agccgttctc accagtccaa gacctggcat gagcggatac atatttgaat
       61 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tacccgaaaa gtgccacctg
      121 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt attacgaggc
      181 cctttcacgc attagatgca tgtcgttaca taacttacgg taaatggccc gcctggctga
      241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      301 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      361 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      421 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      481 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      541 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      601 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      661 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      721 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      781 gaccgatcca gcctccggac tctagcctag gccgcgggac ggataacaat ttcacacagg
      841 aaacagctat gaccattagg cctatttagg tgacactata gaacaagttt gtacaaaaaa
      901 gcaggctggt accggtccgg aattcccggg atatcgcggt ttgcgaactg cgggtggact
      961 gtgtagtgac cggcgtcccg ctgtctcgcc ccgtggcggg tgagcgaggg tgcgtggtgc
     1021 gcggcggcgg cggaacgaac gaacgcggtg cgggcggggc gcccgccgca gggcccatgg
     1081 aacaaaaact tatttctgaa gaagatctgg cctcctgcgc gagcatcgac atcgaggacg
     1141 ccacgcagca cctgcgggac atcctcaagc tggaccggcc cgcgggcggc cccagtgcag
     1201 agagcccacg gccatccagt gcctacaatg gggacctcaa tggacttctg gtcccagacc
     1261 cgctctgctc aggtgatagt acctcagcaa acaagactgg tcttcggacc atgccaccca
     1321 ttaacctgca agagaagcag gtcatctgtc tctcaggaga tgatagctcc acctgcattg
     1381 ggattttggc caaggaggtg gagattgtgg ctagcagtga ctctagcatt tcaagcaagg
     1441 cccggggaag caacaaggtg aaaattcagc ctgtcgccaa gtatgactgg gaacagaagt
     1501 actactatgg caacctgatt gctgtgtcta actccttctt ggcctatgcc attcgggctg
     1561 ccaacaatgg ctctgccatg gtgcgggtga tcagcgtcag cacttcggag cggaccttgc
     1621 tcaagggctt cacaggcagt gtggctgatc tggctttcgc gcacctcaac tctccacagc
     1681 tggcctgcct ggatgaggca ggcaacctgt tcgtgtggcg cttggctctg gttaatggca
     1741 aaattcaaga agagatcttg gtccatattc ggcagccaga gggcacgcca ctgaaccact
     1801 ttcgcaggat catctggtgc cccttcatcc ctgaggagag cgaagactgc tgtgaggaga
     1861 gcagcccaac agtggccctg ctgcatgaag accgggctga ggtgtgggac ctggacatgc
     1921 tccgctccag ccacagtacc tggcctgtgg atgttagcca gatcaagcag ggcttcattg
     1981 tggtaaaagg tcatagcacg tgcctcagtg aaggagccct ctctcctgat gggactgtgc
     2041 tggctactgc gagccacgat ggctatgtca agttctggca gatctacatt gaggggcaag
     2101 atgagccaag gtgtctgcac gagtggaaac ctcatgatgg gcggcccctc tcctgcctcc
     2161 tgttctgtga caaccataag aaacaagacc ctgatgtccc tttctggagg ttccttatta
     2221 ctggtgctga ccagaaccga gagttaaaga tgtggtgtac agtatcctgg acctgcctgc
     2281 agactattcg cttctcccca gatatcttca gctcagtgag tgtgccccct agcctcaagg
     2341 tttgcttgga cctctcagca gaatacctga ttctcagcga tgtgcaacgg aaggtcctct
     2401 atgtgatgga gctgctgcaa aaccaggagg agggccacgc ctgcttcagc tccatctcgg
     2461 agttcctgct cacccaccct gtgctgagct ttggtatcca ggttgtgagt cgctgccggc
     2521 tacggcacac tgaggtgctg cctgccgaag aggaaaatga cagcctgggt gctgatggta
     2581 cccatggagc cggtgccatg gagtctgcgg ccggtgtgct catcaagctc ttttgtgtgc
     2641 atactaaggc actgcaagat gtgcagatcc gcttccagcc acagctgaac cctgatgtgg
     2701 tggccccact gcccacccac actgcccacg aggacttcac atttggagag tctcggcccg
     2761 aactgggctc tgagggcctg gggtcagccg ctcacggctc ccagcctgac ctccgacgaa
     2821 tcgtggagct gcctgcacct gccgacttcc tcagtctgag cagtgagacc aagcccaagt
     2881 tgatgacacc tgacgccttc atgacaccta gcgcctcctt gcagcagatc actgcctctc
     2941 ccagcagcag cagcagcggt agcagcagca gcagcagcag tagcagcagc tcccttacag
     3001 ctgtgtctgc catgagcagc acctcagctg tggacccctc cttgaccagg ccacctgagg
     3061 agctgacctt gagccccaag ctgcagctgg atggcagcct gacaatgagc agcagtggca
     3121 gccttcaggc aagcccgcgt ggcctcctgc ctggcctgct cccagcccca gctgacaaac
     3181 tgactcccaa ggggccgggc caggtgccta ctgccacctc tgcactgtcc ctggagctgc
     3241 aggaagtgga gcccctgggg ctaccccaag cctcccctag ccgcactcgt tcccctgatg
     3301 tcatctcctc agcttccact gccctgtccc aggacatccc tgagattgca tctgaggccc
     3361 tgtcccgtgg ttttggctcc tctgcaccag agggccttga gccagacagt atggcttcag
     3421 ccgcctcggc actgcacctg ctgtccccac ggccccggcc agggcccgag ctcggccccc
     3481 agctcgggct tgatggaggc cctggggatg gagatcggca taataccccc tccctcctgg
     3541 aggcagcctt gacccaggag gcctcgactc ctgacagtca ggtttggccc acagcacctg
     3601 acattactcg tgagacctgc agcaccctgg cagaaagccc caggaatggc cttcaggaaa
     3661 agcacaagag cctggccttc caccgaccac catatcacct gctgcagcaa cgtgacagcc
     3721 aggatgccag tgctgagcaa agtgaccatg atgatgaggt ggccagcctt gcctctgctt
     3781 caggaggctt tggcaccaaa gttcctgctc cacggctgcc tgccaaggac tggaagacca
     3841 agggatcccc tcgaacctca cccaagctca agaggaaaag caagaaggat gatggggatg
     3901 cagccatggg atcccggctc acagagcacc aggtggcaga gccccctgag gactggccag
     3961 cactaatttg gcaacagcag agagagctgg cagagctgcg gcacagccag gaagagctgc
     4021 tgcagcgtct gtgtacccaa ctcgaaggcc tgcagagcac agtcacaggc cacgtagaac
     4081 gtgcccttga gactcggcac gagcaggaac agcggcggct ggagcgagca ctggctgagg
     4141 ggcagcagcg gggagggcag ctgcaggagc agctgacaca acagttgtcc caagcactgt
     4201 cgtcagctgt agctgggcgg ctagagcgca gcatacggga tgagatcaag aagacagtcc
     4261 ctccatgtgt ctcaaggagt ctggagccta tggcaggcca actgagcaac tcagtggcta
     4321 ccaagctcac agctgtggag ggcagcatga aagagaacat ctccaagctg ctcaagtcca
     4381 agaacttgac tgatgccatc gcccgagcag ctgcagacac attacaaggg ccgatgcagg
     4441 ctgcctaccg ggaagccttc cagagtgtgg tgctgccggc ctttgagaag agctgccagg
     4501 ccatgttcca gcaaatcaat gatagcttcc ggctggggac acaggaatac ttgcagcagc
     4561 tagaaagcca catgaagagc cggaaggcac gggaacagga ggccagggag cctgtgctag
     4621 cccagctgcg gggcctggtc agcacactgc agagtgccac tgagcagatg gcagccaccg
     4681 tggccggcag tgttcgtgct gaggtgcagc accagctgca tgtggctgtg ggcagcctgc
     4741 aggagtccat tttagcacag gtacagcgca tcgttaaggg tgaggtgagt gtggcgctca
     4801 aggagcagca ggccgccgtc acctccagca tcatgcaggc catgcgctca gctgctggca
     4861 cacctgtccc ctctgcccac cttgactgcc aggcccagca agcccatatc ctgcagctgc
     4921 tgcagcaggg ccacctcaat caggccttcc agcaggcgct gacagctgct gacctgaacc
     4981 tggtgctgta tgtgtgtgaa actgtggacc cagcccaggt ttttgggcag ccaccctgcc
     5041 cgctctccca gcctgtgctc ctttccctca tccagcagct ggcatctgac cttggcactc
     5101 gaactgacct caagctcagc tacctggaag aggccgtgat gcacctggac cacagtgacc
     5161 ccatcactcg ggaccacatg ggctccgtta tggcccaggt gcgccaaaag ctttttcagt
     5221 tcctgcaggc tgagccacac aactcacttg gcaaagcagc tcggcgtctc agcctcatgc
     5281 tgcatggcct cgtgaccccc agcctccctt agctgctaag cctgccttgc ccaggggtgg
     5341 gatggcactg aaggccagca gacaggccta ggctggggca gggtcacggc tggcctttac
     5401 ctgctcaggc ccccatctct ggggtgtttg ggggtcaggg agcagggagc actggccgtg
     5461 gtctacagcg tgtggtagtc agaaggttta gctgggccca gggcaggtat tgcgcctgct
     5521 tgggttctgc catgcctgga gcatgaccct gagatcgtga caccacttga gtggaatttt
     5581 ccatgttcct ttttacctct aatttggatc tttttgtttt tgaaaaacat tgagaaattc
     5641 aattaaatgc ttttggaata anaaaaaaaa aaaaaaaaaa aaaaaaaaag ggcggccgct
     5701 ctagagtatc cctcgagggg cccaagctta cgcgtaccca gctttcttgt acaaagtggt
     5761 ccctatagtg agtcgtatta taagctaggc actggccgtc gttttacaac gtcgtgactg
     5821 ggaaaactgc tagcttggga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt
     5881 ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata
     5941 atgtgttaaa ctagctgcat atgcttgctg cttgagagtt ttgcttactg agtatgattt
     6001 atgaaaatat tatacacagg agctagtgat tctaattgtt tgtgtatttt agattcacag
     6061 tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca taatcagcca
     6121 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     6181 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     6241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcgatc ctgcattaat
     6361 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6421 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6481 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6541 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6601 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6661 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6721 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6781 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6841 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6901 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6961 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7021 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7081 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7141 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7201 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg ccataacttc
     7261 gtatagcata cattatacga agttatggca tgagattatc aaaaaggatc ttcacctaga
     7321 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7381 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7441 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7501 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7561 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7621 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7681 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7741 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7801 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7861 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7921 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7981 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8041 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8101 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8161 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8221 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8281 atcagggtta ttgtctcatg ccaggggtgg gcacacatat ttgataccag cgatccctac
     8341 acagcacata attcaatgcg acttccctct atcgcacatc ttagaccttt attctccctc
     8401 cagcacacat cgaag
//
