LOCUS       sequence-170526-        6118 bp    DNA     circular SYN 14-APR-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 6118)
  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 6118)
  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: HA-Beclin 1
FEATURES             Location/Qualifiers
     source          1..6118
                     /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      2418..2427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2431..2565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2468..2487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2522..2541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2634..2991
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2634..2825
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2657..2677)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2842..2977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3011..3028)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3011..3027)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3020..3042)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3240..3695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3327..3346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3537..3558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3712..3731)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3831..3853
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3891..3909)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3977..4081
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4082..4150,4151..4942)
                     /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(4300..4319)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5113..5701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5602..5621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5855..5872
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5989..6010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6025..6030,6031..6048,6049..6055)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6063..6079
                     /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     6068..6090
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6087..6103
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6087..6103
                     /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 catggaaggg tctaagacgt ccaacaacag caccatgcag gtgagcttcg
     1081 tgtgccagcg ctgcagccag cccctgaaac tggacacgag tttcaagatc ctggaccgtg
     1141 tcaccatcca ggaactcaca gctccattac ttaccacagc ccaggcgaaa ccaggagaga
     1201 cccaggagga agagactaac tcaggagagg agccatttat tgaaactcct cgccaggatg
     1261 gtgtctctcg cagattcatc cccccagcca ggatgatgtc cacagaaagt gccaacagct
     1321 tcactctgat tggggaggca tctgatggcg gcaccatgga gaacctcagc cgaagactga
     1381 aggtcactgg ggaccttttt gacatcatgt cgggccagac agatgtggat cacccactct
     1441 gtgaggaatg cacagatact cttttagacc agctggacac tcagctcaac gtcactgaaa
     1501 atgagtgtca gaactacaaa cgctgtttgg agatcttaga gcaaatgaat gaggatgaca
     1561 gtgaacagtt acagatggag ctaaaggagc tggcactaga ggaggagagg ctgatccagg
     1621 agctggaaga cgtggaaaag aaccgcaaga tagtggcaga aaatctcgag aaggtccagg
     1681 ctgaggctga gagactggat caggaggaag ctcagtatca gagagaatac agtgaattta
     1741 aacgacagca gctggagctg gatgatgagc tgaagagtgt tgaaaaccag atgcgttatg
     1801 cccagacgca gctggataag ctgaagaaaa ccaacgtctt taatgcaacc ttccacatct
     1861 ggcacagtgg acagtttggc acaatcaata acttcaggct gggtcgcctg cccagtgttc
     1921 ccgtggaatg gaatgagatt aatgctgctt ggggccagac tgtgttgctg ctccatgctc
     1981 tggccaataa gatgggtctg aaatttcaga gataccgact tgttccttac ggaaaccatt
     2041 catatctgga gtctctgaca gacaaatcta aggagctgcc gttatactgt tctggggggt
     2101 tgcggttttt ctgggacaac aagtttgacc atgcaatggt ggctttcctg gactgtgtgc
     2161 agcagttcaa agaagaggtt gagaaaggcg agacacgttt ttgtcttccc tacaggatgg
     2221 atgtggagaa aggcaagatt gaagacacag gaggcagtgg cggctcctat tccatcaaaa
     2281 cccagtttaa ctctgaggag cagtggacaa aagctctcaa gttcatgctg acgaatctta
     2341 agtggggtct tgcttgggtg tcctcacaat tttataacaa atgagcggcc gctaagtaag
     2401 taaggatccc cagcttggcc gccatggccc aacttgttta ttgcagctta taatggttac
     2461 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2521 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttaattcggc
     2581 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     2641 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2821 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2881 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     2941 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctgtta
     3001 acagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3061 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3121 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     3181 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3241 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3301 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3361 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3421 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     3481 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3541 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     3601 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3661 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3721 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3781 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3841 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     3901 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3961 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4021 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4081 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4141 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4201 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4261 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4321 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4381 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4441 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4501 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4561 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4621 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4681 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4741 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4801 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4861 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4921 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4981 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5041 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5101 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5161 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5221 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5281 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5341 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5401 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5461 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5521 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5581 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5641 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5701 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5761 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5821 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5881 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     5941 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6001 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     6061 aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta cgaattaa
//
