LOCUS       sequence-170519-        5299 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 5299)
  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 5299)
  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 (1-181)
FEATURES             Location/Qualifiers
     source          1..5299
                     /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      1599..1608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1612..1746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1649..1668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1703..1722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1815..2172
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1815..2006
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1838..1858)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2023..2158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2192..2209)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2192..2208)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2201..2223)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2421..2876
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2508..2527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2718..2739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2893..2912)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3012..3034
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3072..3090)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3158..3262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3263..3331,3332..4123)
                     /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(3481..3500)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4294..4882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4783..4802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5036..5053
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5170..5191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5206..5211,5212..5229,5230..5236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5244..5260
                     /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     5249..5271
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5268..5284
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5268..5284
                     /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 ctggtggact 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 agaccgcggc cgctaagtaa gtaaggatcc ccagcttggc cgccatggcc caacttgttt
     1621 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1681 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1741 tggatcggga attaattcgg cgcagcacca tggcctgaaa taacctctga aagaggaact
     1801 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     1861 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1921 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1981 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2041 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2101 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2161 ggcttttgca aaaagctgtt aacagcttgg cactggccgt cgttttacaa cgtcgtgact
     2221 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2281 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2341 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2401 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2461 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2521 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2581 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     2641 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2701 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2761 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2821 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     2881 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2941 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3001 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3061 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3121 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3181 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3241 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3301 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3361 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3421 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3481 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3541 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3601 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3661 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3721 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3781 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3841 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3901 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3961 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4021 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4081 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4141 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4201 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4261 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4321 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4381 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4441 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4501 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4561 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4621 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4681 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4741 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4801 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4861 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4921 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4981 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5041 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5101 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5161 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     5221 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     5281 tgacatgatt acgaattaa
//
