LOCUS       sequence-170503-        6865 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 6865)
  AUTHORS   Kim YM, Jung CH, Seo M, Kim EK, Park JM, Bae SS, Kim DH
  TITLE     mTORC1 phosphorylates UVRAG to negatively regulate autophagosome and
            endosome maturation.
  JOURNAL   Mol Cell. 2015 Jan 22;57(2):207-18. doi: 
            10.1016/j.molcel.2014.11.013. Epub 2014 Dec 18.
  PUBMED    25533187
REFERENCE   2  (bases 1 to 6865)
  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"; doi: 
            "10.1016/j.molcel.2014.11.013"; journalName: "Mol Cell"; date: 
            "2015-01-22- 22"; volume: "57"; issue: "2"; pages: "207-18"
COMMENT     Sequence Label: HA-UVRAG
FEATURES             Location/Qualifiers
     source          1..6865
                     /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      3165..3174
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3178..3312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3215..3234)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3269..3288
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3381..3738
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3381..3572
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3404..3424)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3589..3724
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3758..3775)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3758..3774)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3767..3789)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3987..4442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4074..4093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4284..4305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4459..4478)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4578..4600
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4638..4656)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4724..4828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4829..4897,4898..5689)
                     /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(5047..5066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5860..6448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6349..6368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6602..6619
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6736..6757
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6772..6777,6778..6795,6796..6802)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6810..6826
                     /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     6815..6837
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6834..6850
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6834..6850
                     /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 catgagcgcc tccgcgtcgg tcgggggcca ggtcccccag ccacccccgg
     1081 gcccggccgc tgctctgcct cccggttctg ccgcgcgggc cctgcatgtg gagctgccgt
     1141 ctcagcagcg gcgtcttcga catcttcgga acattgctgc ccggaacatt gttaatagaa
     1201 atggccatca gctccttgat acctacttta cacttcactt gtgtagtact gaaaagatat
     1261 ataaagaatt ttatagaagt gaagtgatta agaattcctt gaatcccacg tggcgaagtc
     1321 tcgattttgg aattatgcca gaccgtcttg atacatctgt gtcttgtttc gtggtgaaga
     1381 tatggggtgg aaaggagaac atctaccagc tgttgattga atggaaagtc tgtttggatg
     1441 ggctgaaata cttgggtcag cagattcatg cccgaaacca aaatgaaata atttttgggc
     1501 tgaatgatgg atactatggt gctccatttg aacataaggg ttattcaaat gctcagaaga
     1561 ctattcttct gcaggtggat cagaactgtg ttcgcaattc ttacgatgtc ttctctttgc
     1621 tacggcttca tagagcccag tgtgcaatta aacagactca ggtaactgtt cagaaaattg
     1681 gaaaggaaat tgaagaaaaa ctaagactca catctacaag caatgaactg aaaaaaaaaa
     1741 gtgaatgcct gcagttaaaa attttggtgc ttcagaatga actggaacgg cagaagaaag
     1801 ctttgggacg ggaggtggca ttactgcata agcaacaaat tgcattacaa gacaaaggaa
     1861 gtgcattttc agctgagcac ctcaaacttc aactccagaa ggaatcccta aatgagctga
     1921 ggaaggagtg cactgcaaaa agagaactct tcttgaagac taatgctcag ttgacaattc
     1981 gttgcaggca gttactctct gagctttcct acatttaccc tattgatttg aatgaacata
     2041 aggattactt tgtatgcggt gtcaagttgc ctaattctga ggacttccaa gcaaaagatg
     2101 atggaagcat tgctgttgcc cttggttata ctgcacatct ggtctccatg atttcctttt
     2161 tcctacaagt gcccctcaga tatcctataa ttcataaggg gtctagatca acaatcaaag
     2221 acaatatcaa tgacaaactg acggaaaagg agagagagtt tccactgtat ccaaaaggag
     2281 gggagaagtt gcagtttgat tatggtgtct atcttctgaa caaaaatata gcacagctaa
     2341 gatatcaaca tggactaggg actccagact tgcggcaaac ccttcccaac ctgaaaaact
     2401 tcatggagca tggactaatg gtcaggtgtg acagacatca cacctccagt gcaatccctg
     2461 ttcctaagag acaaagctcc atatttgggg gtgcagatgt aggcttctct ggggggatcc
     2521 cttcaccaga caaaggacat cgaaaacggg ccagctctga gaatgagaga cttcagtaca
     2581 aaacccctcc tcccagttac aactcagcat tagcccagcc tgtgaccacc gtcccctcca
     2641 tgggagagac cgagagaaag ataacatctc tatcctcctc cttggatacc tccttggact
     2701 tctccaaaga aaacaagaaa aaaggagagg atctagttgg cagcttaaac ggaggccacg
     2761 cgaatgtgca ccctagccaa gaacaaggag aagccctctc cgggcaccgg gccacagtca
     2821 atggcactct cctacccagc gagcaggccg ggtccgccag tgtccagctt ccaggcgagt
     2881 tccacccagt ctcagaagct gagctctgct gtactgtgga gcaagcagaa gaaatcatcg
     2941 ggctggaagc cacaggtttc gcctcaggtg atcagctaga agcatttaac tgcatcccag
     3001 tggacagtgc tgtggcagta gagtgtgacg aacaagttct gggagaattt gaagagttct
     3061 cccgaaggat ctatgcactg aatgaaaacg tatccagctt ccgccggccg cgcaggagtt
     3121 ccgataagtg agcggccgct aagtaagtaa ggatccccag cttggccgcc atggcccaac
     3181 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3241 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3301 catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac ctctgaaaga
     3361 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3421 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3481 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3541 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3601 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3661 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3721 ggcctaggct tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc
     3781 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     3841 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3901 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3961 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     4021 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4081 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4141 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4201 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4261 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4321 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4381 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     4441 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4501 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4561 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4621 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4681 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4741 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4801 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4861 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4921 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4981 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5041 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5101 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5161 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5221 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5281 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5341 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5401 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5461 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5521 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5581 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5641 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5701 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5761 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5821 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5881 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5941 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6001 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6061 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6121 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6181 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6241 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6301 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6361 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6421 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6481 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6541 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6601 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6661 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6721 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6781 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6841 cagctatgac atgattacga attaa
//
