LOCUS       sequence-170523-        6238 bp    DNA     circular SYN 15-DEC-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 6238)
  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 6238)
  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: 
            "Autophagy."; date: "2016"; volume: "12(3):547-64. doi"; pages: " 
            10.1080/15548627.2016.1140293"
COMMENT     Sequence Label: myc-Atg14 S29A
FEATURES             Location/Qualifiers
     source          1..6238
                     /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      2538..2547
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2551..2685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2588..2607)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2642..2661
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2754..3111
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2754..2945
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2777..2797)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2962..3097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3131..3148)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3131..3147)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3140..3162)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3360..3815
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3447..3466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3657..3678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3832..3851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3951..3973
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4011..4029)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4097..4201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4202..4270,4271..5062)
                     /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(4420..4439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5233..5821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5722..5741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5975..5992
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6109..6130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6145..6150,6151..6168,6169..6175)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6183..6199
                     /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     6188..6210
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6207..6223
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6207..6223
                     /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 cgaccatggc gtctcccagt gggaagggag cccgggcgct ggaggctcct ggctgcgggc
     1081 cccggccgct cgcccgggac ctggtggacg ccgtggacga tgcggagggg ctgtacgtgg
     1141 ctgtggagcg ctgcccgctg tgcaacacta cccgccggcg gctgacctgc gccaaatgcg
     1201 ttcagagcgg cgatttcgtc tacttcgacg gccgcgaccg ggagaggttt atcgacaaga
     1261 aggaaaggtt aagccgactt aagagcaagc aagaagaatt tcagaaagaa gtgttaaaag
     1321 ctatggaagg aaaatggata acagatcagt tgagatggaa aataatgtcc tgcaagatga
     1381 ggattgaaca gttaaaacaa acaatatgta aaggaaatga agaaatggag aaaaattctg
     1441 aaggccttct caaaaccaag gaaaagaatc agaagcttta cagtcgagca caacggcacc
     1501 aagagaaaaa ggagaagatt cagaggcata atcgcaaact tggtgacctg gtagaaaaaa
     1561 agaccattga cttaagaagt cattatgagc gtctggcaaa tcttcgacga tcccatatat
     1621 tagagctcac ctctgtcatt tttccaatcg aggaagtaaa gacgggtgtg agagaccccg
     1681 cagatgtgtc ttcagagagt gacagtgcca tgacctccag cactgtgagc aagcttgctg
     1741 aagcccggag gacaacttac ctctcaggac gatgggtctg tgacgatcac aacggagaca
     1801 ccagcattag cattacaggg ccttggatta gcctccctaa caatggggac tactctgcct
     1861 actacagctg ggtggaggag aagaaaacaa cccaggggcc tgacatggag cagagtaacc
     1921 ctgcctacac catcagtgct gcgctgtgct atgcaactca gctggtcaac attctgtctc
     1981 atatacttga tgtaaatctt cccaaaaagc tctgcaacag tgaattttgt ggcgaaaatc
     2041 taagcaagca gaaatttact cgagcagtga agaaactgaa tgcaaatatt ctttaccttt
     2101 gtttttctca gcatgtaaat ttagatcaat tacaaccact gcataccctc aggaatctaa
     2161 tgtacctggt cagtccaagc tctgaacacc taggcaggtc agggcccttt gaagtacgag
     2221 cagaccttga ggagtccatg gaatttgtgg atcccggagt tgctggagaa tcagatgaga
     2281 gcggagatga gcgcgtcagc gatgaagaaa ccgacctggg cacagactgg gagaacttgc
     2341 ctagtccccg gttttgtgat atcccttccc agtctgtgga agtctcccag agtcagagca
     2401 cccaggcgtc cccacccatc gcgagcagca gtgcaggtgg gatgatctcc tctgcagcag
     2461 cctcggtgac ctcctggttt aaagcttaca ctggacaccg ttaagcggcc gctaagtaag
     2521 taaggatccc cagcttggcc gccatggccc aacttgttta ttgcagctta taatggttac
     2581 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2641 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttaattcggc
     2701 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     2761 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2941 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3001 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3061 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctgtta
     3121 acagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3181 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3241 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     3301 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3361 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3421 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3481 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3541 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     3601 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3661 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     3721 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3781 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3841 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3901 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3961 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4021 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4081 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4141 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4201 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4261 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4321 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4381 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4441 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4501 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4561 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4621 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4681 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4741 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4801 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4861 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4921 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4981 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5041 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5101 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5161 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5221 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5281 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5341 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5401 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5461 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5521 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5581 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5641 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5701 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5761 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5821 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5881 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5941 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6001 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6061 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6121 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     6181 aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta cgaattaa
//
