LOCUS       sequence-170527-        7429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal HA-tagged protein expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7429)
  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 7429)
  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: HA-Vps34
FEATURES             Location/Qualifiers
     source          1..7429
                     /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             975..1001
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3729..3738
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3742..3876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3779..3798)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3833..3852
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3945..4302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3945..4136
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3968..3988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4153..4288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4322..4339)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4322..4338)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4331..4353)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4551..5006
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4638..4657)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4848..4869
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5023..5042)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5142..5164
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5202..5220)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5288..5392
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5393..5461,5462..6253)
                     /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(5611..5630)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6424..7012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6913..6932
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7166..7183
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7300..7321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7336..7341,7342..7359,7360..7366)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7374..7390
                     /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     7379..7401
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7398..7414
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7398..7414
                     /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 gggctacccc tatgatgtgc ctgactacgc agatctcaat ggtggtggtg
     1021 gtgggtcgac catgggggaa gcagagaagt ttcactacat ctatagttgt gacctggata
     1081 tcaacgtcca gcttaagata ggaagcttgg aagggaagag agaacaaaag agttataaag
     1141 ctgtcctgga agacccaatg ttgaagttct caggactata tcaagagaca tgctctgatc
     1201 tttatgttac ttgtcaagtt tttgcagaag ggaagccttt ggccttgcca gtgagaacat
     1261 cctacaaagc atttagtaca agatggaact ggaatgaatg gctgaaacta ccagtaaaat
     1321 accctgacct gcccaggaat gcccaagtgg ccctcaccat atgggatgtg tatggtcccg
     1381 gaaaagcagt gcctgtagga ggaacaacgg tttcgctctt tggaaaatac ggcatgtttc
     1441 gccaagggat gcatgacttg aaagtctggc ctaatgtaga agcagatgga tcagaaccca
     1501 caaaaactcc tggcagaaca agtagcactc tctcagaaga tcagatgagc cgtcttgcca
     1561 agctcaccaa agctcatcga caaggacaca tggtgaaagt agattggctg gatagattga
     1621 catttagaga aatagaaatg ataaatgaga gtgaaaaacg aagttctaat ttcatgtacc
     1681 tgatggttga atttcgatgt gtcaagtgtg atgataagga atatggtatt gtttattatg
     1741 aaaaggacgg tgatgaatca tctccaattt taacaagttt tgaattagtg aaagttcctg
     1801 acccccagat gtctatggag aatttagttg agagcaaaca ccacaagctt gcccggagtt
     1861 taagaagtgg accttctgac cacgatctga aacccaatgc tgccacgaga gatcagttaa
     1921 atattattgt gagttatcca ccaaccaagc aacttacata tgaagaacaa gatcttgttt
     1981 ggaagtttag atattatctt acgaatcaag aaaaagcctt gacaaaattc ttgaaatgtg
     2041 ttaattggga tctacctcaa gaggccaaac aggccttgga acttctggga aaatggaagc
     2101 cgatggatgt agaggactcc ttggagctgt tatcctctca ttacaccaac ccaactgtga
     2161 ggcgttatgc tgttgcccgg ttgcgacagg ccgatgatga ggatttgttg atgtacctat
     2221 tacaattggt ccaggctctc aaatatgaaa attttgatga tataaagaat ggattggaac
     2281 ctaccaagaa ggatagtcag agttcagtgt cagaaaatgt gtcaaattct ggaataaatt
     2341 ctgcagaaat agatagctcc caaattataa ccagccccct tccttcagtc tcttcacctc
     2401 ctcctgcatc aaaaacaaaa gaagttccag atggcgaaaa tctggaacaa gatctctgta
     2461 ccttcttgat atcgagagcc tgcaaaaact caacactggc taattattta tactggtatg
     2521 tgatagtgga atgtgaagat caagatactc agcagagaga tccaaagacc catgagatgt
     2581 acttgaacgt aatgagaaga ttcagccaag cattgttgaa gggtgataag tctgtcagag
     2641 ttatgcgttc tttgctggct gcacaacaga catttgtaga tcggttggtg catctaatga
     2701 aggcagtaca acgcgaaagt ggaaatcgta agaaaaagaa tgagagacta caggcattgc
     2761 ttggagataa tgaaaagatg aatttgtcag atgtggaact tatcccgttg cctttagaac
     2821 cccaagtgaa aattagagga ataattccgg aaacagctac actgtttaaa agtgccctta
     2881 tgcctgcaca gttgtttttt aagacggaag atggaggcaa atatccagtt atatttaagc
     2941 atggagatga tttacgtcaa gatcaactta ttcttcaaat catttcactc atggacaagc
     3001 tgttacggaa agaaaatctg gacttgaaat tgacacctta taaggtgtta gccaccagta
     3061 caaaacatgg cttcatgcag tttatccagt cagttcctgt ggctgaagtt cttgatacag
     3121 agggaagcat tcagaacttt tttagaaaat atgcaccaag tgagaatggg ccaaatggga
     3181 ttagtgctga ggtcatggac acttacgtta aaagctgtgc tggatattgc gtgatcacct
     3241 atatacttgg agttggagac aggcacctgg ataacctttt gctaacaaaa acaggcaaac
     3301 tcttccacat agactttgga tatattttgg gtcgggatcc aaagcctctt cctccaccaa
     3361 tgaagctgaa taaagaaatg gtagaaggaa tggggggcac acagagtgag cagtaccaag
     3421 agttccgtaa acagtgttac acggctttcc tccacctgcg aaggtattct aatctgattt
     3481 tgaacttgtt ttccttgatg gttgatgcaa acattccaga tattgcactt gaaccagata
     3541 aaactgtgaa aaaggttcag gataaattcc gcttagacct gtcggatgaa gaggctgtgc
     3601 attacatgca gagtctgatt gatgagagtg tccatgctct ttttgctgca gtggtggaac
     3661 agattcacaa gtttgcccag tactggagaa aatgagcggc cgctaagtaa gtaaggatcc
     3721 ccagcttggc cgccatggcc caacttgttt attgcagctt ataatggtta caaataaagc
     3781 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3841 tccaaactca tcaatgtatc ttatcatgtc tggatcggga attaattcgg cgcagcacca
     3901 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     3961 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4021 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4081 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4141 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4201 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4261 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctgtt aacagcttgg
     4321 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4381 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4441 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     4501 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4561 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4621 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4681 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4741 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4801 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4861 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     4921 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4981 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     5041 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5101 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5161 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5221 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5281 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5341 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5401 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5461 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5521 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5581 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5641 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5701 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5761 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5821 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5881 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5941 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6001 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6061 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6121 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6181 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6241 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6301 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6361 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6421 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6481 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6541 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6601 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6661 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6721 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6781 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6841 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6901 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6961 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7021 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7081 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7141 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7201 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7261 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7321 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7381 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaattaa
//
