LOCUS       sequence-170504-        6859 bp    DNA     circular SYN 15-APR-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 6859)
  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 6859)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: myc-UVRAG S498D
FEATURES             Location/Qualifiers
     source          1..6859
                     /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      3159..3168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3172..3306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3209..3228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3263..3282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3375..3732
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3375..3566
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3398..3418)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3583..3718
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3752..3769)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3752..3768)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3761..3783)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3981..4436
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4068..4087)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4278..4299
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4453..4472)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4572..4594
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4632..4650)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4718..4822
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4823..4891,4892..5683)
                     /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(5041..5060)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5854..6442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6343..6362
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6596..6613
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6730..6751
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6766..6771,6772..6789,6790..6796)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6804..6820
                     /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     6809..6831
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6828..6844
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6828..6844
                     /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 cgaccatgag cgcctccgcg tcggtcgggg gccaggtccc ccagccaccc ccgggcccgg
     1081 ccgctgctct gcctcccggt tctgccgcgc gggccctgca tgtggagctg ccgtctcagc
     1141 agcggcgtct tcgacatctt cggaacattg ctgcccggaa cattgttaat agaaatggcc
     1201 atcagctcct tgatacctac tttacacttc acttgtgtag tactgaaaag atatataaag
     1261 aattttatag aagtgaagtg attaagaatt ccttgaatcc cacgtggcga agtctcgatt
     1321 ttggaattat gccagaccgt cttgatacat ctgtgtcttg tttcgtggtg aagatatggg
     1381 gtggaaagga gaacatctac cagctgttga ttgaatggaa agtctgtttg gatgggctga
     1441 aatacttggg tcagcagatt catgcccgaa accaaaatga aataattttt gggctgaatg
     1501 atggatacta tggtgctcca tttgaacata agggttattc aaatgctcag aagactattc
     1561 ttctgcaggt ggatcagaac tgtgttcgca attcttacga tgtcttctct ttgctacggc
     1621 ttcatagagc ccagtgtgca attaaacaga ctcaggtaac tgttcagaaa attggaaagg
     1681 aaattgaaga aaaactaaga ctcacatcta caagcaatga actgaaaaaa aaaagtgaat
     1741 gcctgcagtt aaaaattttg gtgcttcaga atgaactgga acggcagaag aaagctttgg
     1801 gacgggaggt ggcattactg cataagcaac aaattgcatt acaagacaaa ggaagtgcat
     1861 tttcagctga gcacctcaaa cttcaactcc agaaggaatc cctaaatgag ctgaggaagg
     1921 agtgcactgc aaaaagagaa ctcttcttga agactaatgc tcagttgaca attcgttgca
     1981 ggcagttact ctctgagctt tcctacattt accctattga tttgaatgaa cataaggatt
     2041 actttgtatg cggtgtcaag ttgcctaatt ctgaggactt ccaagcaaaa gatgatggaa
     2101 gcattgctgt tgcccttggt tatactgcac atctggtctc catgatttcc tttttcctac
     2161 aagtgcccct cagatatcct ataattcata aggggtctag atcaacaatc aaagacaata
     2221 tcaatgacaa actgacggaa aaggagagag agtttccact gtatccaaaa ggaggggaga
     2281 agttgcagtt tgattatggt gtctatcttc tgaacaaaaa tatagcacag ctaagatatc
     2341 aacatggact agggactcca gacttgcggc aaacccttcc caacctgaaa aacttcatgg
     2401 agcatggact aatggtcagg tgtgacagac atcacacctc cagtgcaatc cctgttccta
     2461 agagacaaag ctccatattt gggggtgcag atgtaggctt ctctgggggg atccctgacc
     2521 cagacaaagg acatcgaaaa cgggccagct ctgagaatga gagacttcag tacaaaaccc
     2581 ctcctcccag ttacaactca gcattagccc agcctgtgac caccgtcccc tccatgggag
     2641 agaccgagag aaagataaca tctctatcct cctccttgga tacctccttg gacttctcca
     2701 aagaaaacaa gaaaaaagga gaggatctag ttggcagctt aaacggaggc cacgcgaatg
     2761 tgcaccctag ccaagaacaa ggagaagccc tctccgggca ccgggccaca gtcaatggca
     2821 ctctcctacc cagcgagcag gccgggtccg ccagtgtcca gcttccaggc gagttccacc
     2881 cagtctcaga agctgagctc tgctgtactg tggagcaagc agaagaaatc atcgggctgg
     2941 aagccacagg tttcgcctca ggtgatcagc tagaagcatt taactgcatc ccagtggaca
     3001 gtgctgtggc agtagagtgt gacgaacaag ttctgggaga atttgaagag ttctcccgaa
     3061 ggatctatgc actgaatgaa aacgtatcca gcttccgccg gccgcgcagg agttccgata
     3121 agtgagcggc cgctaagtaa gtaaggatcc ccagcttggc cgccatggcc caacttgttt
     3181 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3241 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3301 tggatcggga attaattcgg cgcagcacca tggcctgaaa taacctctga aagaggaact
     3361 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3481 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3541 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3601 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3661 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3721 ggcttttgca aaaagctgtt aacagcttgg cactggccgt cgttttacaa cgtcgtgact
     3781 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3841 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3901 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3961 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4021 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4081 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4141 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4201 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4261 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4321 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4381 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4441 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4501 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4561 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4621 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4681 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4741 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4801 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4861 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4921 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4981 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5041 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5101 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5161 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5221 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5281 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5341 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5401 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5461 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5521 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5581 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5641 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5701 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5761 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5821 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5881 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5941 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6001 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6061 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6121 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6181 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6241 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6301 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6361 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6421 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6481 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6541 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6601 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6661 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     6721 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     6781 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     6841 tgacatgatt acgaattaa
//
