LOCUS       sequence-170521-        5461 bp    DNA     circular SYN 14-APR-2025
DEFINITION  N-terminal myc-tagged protein expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5461)
  AUTHORS   Park JM, Jung CH, Seo M, Otto NM, Grunwald D, Kim KH, Moriarity B, 
            Kim YM, Starker C, Nho RS, Voytas D, Kim DH
  TITLE     The ULK1 complex mediates MTORC1 signaling to the autophagy 
            initiation machinery via binding and phosphorylating ATG14.
  JOURNAL   Autophagy. 2016;12(3):547-64. doi: 10.1080/15548627.2016.1140293.
  PUBMED    27046250
REFERENCE   2  (bases 1 to 5461)
  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: 
            "Autophagy."; date: "2016"; volume: "12(3):547-64. doi"; pages: " 
            10.1080/15548627.2016.1140293"
COMMENT     Sequence Label: myc-Atg14 (181-413)
FEATURES             Location/Qualifiers
     source          1..5461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      963..972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             972..1001
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1761..1770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1774..1908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1811..1830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1865..1884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1977..2334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1977..2168
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2000..2020)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2185..2320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2354..2371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2354..2370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2363..2385)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2583..3038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2670..2689)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2880..2901
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3055..3074)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3174..3196
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3234..3252)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3320..3424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3425..3493,3494..4285)
                     /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     complement(3643..3662)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4456..5044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4945..4964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5198..5215
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5332..5353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5368..5373,5374..5391,5392..5398)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5406..5422
                     /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)."
     primer_bind     5411..5433
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5430..5446
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5430..5446
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc aaggccattc
      961 gcgccaccat ggagcaaaag ctcatttctg aggaagatct caatggtggt ggtggtgggt
     1021 cgaccaccat tgacttaaga agtcattatg agcgtctggc aaatcttcga cgatcccata
     1081 tattagagct cacctctgtc atttttccaa tcgaggaagt aaagacgggt gtgagagacc
     1141 ccgcagatgt gtcttcagag agtgacagtg ccatgacctc cagcactgtg agcaagcttg
     1201 ctgaagcccg gaggacaact tacctctcag gacgatgggt ctgtgacgat cacaacggag
     1261 acaccagcat tagcattaca gggccttgga ttagcctccc taacaatggg gactactctg
     1321 cctactacag ctgggtggag gagaagaaaa caacccaggg gcctgacatg gagcagagta
     1381 accctgccta caccatcagt gctgcgctgt gctatgcaac tcagctggtc aacattctgt
     1441 ctcatatact tgatgtaaat cttcccaaaa agctctgcaa cagtgaattt tgtggcgaaa
     1501 atctaagcaa gcagaaattt actcgagcag tgaagaaact gaatgcaaat attctttacc
     1561 tttgtttttc tcagcatgta aatttagatc aattacaacc actgcatacc ctcaggaatc
     1621 taatgtacct ggtcagtcca agctctgaac acctaggcag gtcagggccc tttgaagtac
     1681 gagcagacct tgaggagtcc atggaatttg tggatcccgg agttgctgcg gccgctaagt
     1741 aagtaaggat ccccagcttg gccgccatgg cccaacttgt ttattgcagc ttataatggt
     1801 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1861 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcgg gaattaattc
     1921 ggcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     1981 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2041 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2101 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2161 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2221 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2281 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctg
     2341 ttaacagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2401 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2461 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     2521 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2581 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2641 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2701 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2761 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2821 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2881 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt
     2941 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3001 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3061 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3121 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3181 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3241 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3301 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3361 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3421 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3481 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3541 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3601 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3661 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3721 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3781 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3841 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3901 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3961 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4021 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4081 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4141 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4201 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4261 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4321 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4381 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4441 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4501 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4561 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4621 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4681 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4741 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4801 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4861 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4921 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4981 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5041 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5101 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5161 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5221 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     5281 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     5341 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     5401 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaatta
     5461 a
//
