LOCUS       sequence-221630-        5769 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5769)
  AUTHORS   Miller DW, Ahmad R, Hague S, Baptista MJ, Canet-Aviles R, McLendon 
            C, Carter DM, Zhu PP, Stadler J, Chandran J, Klinefelter GR, 
            Blackstone C, Cookson MR
  TITLE     L166P mutant DJ-1, causative for recessive Parkinson's disease, is 
            degraded through the ubiquitin-proteasome system.
  JOURNAL   J Biol Chem. 2003 Sep 19. 278(38):36588-95.
  PUBMED    12851414
REFERENCE   2  (bases 1 to 5769)
  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: "J Biol 
            Chem. 2003 Sep 19. 278(38):36588-95."
COMMENT     Sequence Label: pGW1-Myc-DJ1-WT
FEATURES             Location/Qualifiers
     source          1..5769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          720..1546
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     regulatory      1570..1579
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1579..1608
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2357..2491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2394..2413)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2448..2467
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2514..2530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2514..2530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2527..2549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2538..2554
                     /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)."
     promoter        complement(join(2562..2568,2569..2586,2587..2592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2744..2761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2915..3503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2995..3014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3674..4465,4466..4534))
                     /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     4297..4316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     4744..4762
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4800..4822)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4922..4941
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5173..5195
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5187..5204
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5188..5204
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcttatgca tgctatactg
      781 tttttggctt ggggtctata cacccccgct tcctcatgtt ataggtgatg gtatagctta
      841 gcctataggt gtgggttatt gaccattatt gaccactccc ctattggtga cgatactttc
      901 cattactaat ccataacatg gctctttgcc acaactctct ttattggcta tatgccaata
      961 cactgtcctt cagagactga cacggactct gtatttttac aggatggggt ctcatttatt
     1021 atttacaaat tcacatatac aacaccaccg tccccagtgc ccgcagtttt tattaaacat
     1081 aacgtgggat ctccacgcga atctcgggta cgtgttccgg acatgggctc ttctccggta
     1141 gcggcggagc ttctacatcc gagccctgct cccatgcctc cagcgactca tggtcgctcg
     1201 gcagctcctt gctcctaaca gtggaggcca gacttaggca cagcacgatg cccaccacca
     1261 ccagtgtgcc gcacaaggcc gtggcggtag ggtatgtgtc tgaaaatgag ctcggggagc
     1321 gggcttgcac cgctgacgca tttggaagac ttaaggcagc ggcagaagaa gatgcaggca
     1381 gctgagttgt tgtgttctga taagagtcag aggtaactcc cgttgcggtg ctgttaacgg
     1441 tggagggcag tgtagtctga gcagtactcg ttgctgccgc gcgcgccacc agacataata
     1501 gctgacagac taacagactg ttcctttcca tgggtctttt ctgcagtcac cgtccttgac
     1561 acgaagcttg ccaccatgga gcaaaagctg atatctgaag aggacttggg cgcgcctgcc
     1621 cgggataggt acctcgcgag atctagagtc gacctgcagg atatcgaatt catggcttcc
     1681 aaaagagctc tggtcatcct ggctaaagga gcagaggaaa tggagacggt catccctgta
     1741 gatgtcatga ggcgagctgg gattaaggtc accgttgcag gcctggctgg aaaagaccca
     1801 gtacagtgta gccgtgatgt ggtcatttgt cctgatgcca gccttgaaga tgcaaaaaaa
     1861 gagggaccat atgatgtggt ggttctacca ggaggtaatc tgggcgcaca gaatttatct
     1921 gagtctgctg ctgtgaagga gatactgaag gagcaggaaa accggaaggg cctgatagcc
     1981 gccatctgtg caggtcctac tgctctgttg gctcatgaaa taggttttgg aagtaaagtt
     2041 acaacacacc ctcttgctaa agacaaaatg atgaatggag gtcattacac ctactctgag
     2101 aatcgtgtgg aaaaagacgg cctgattctt acaagccggg ggcctgggac cagcttcgag
     2161 tttgcgcttg caattgttga agccctgaat ggcaaggagg tggcggctca agtgaaggct
     2221 ccacttgttc ttaaagacta ggaattcatt gatcataatc agccatacca catttgtaga
     2281 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2341 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2401 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2461 actcatcaat gtatcttatc atgtctggat ccgtaccgag ctcgcgtaat catgtcatag
     2521 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2581 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tacattaatt gcgttgcgct
     2641 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2701 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2761 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2821 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2881 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2941 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3001 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3061 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3121 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3181 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3241 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3301 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3361 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3421 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3481 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3541 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3601 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3661 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3721 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3781 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3841 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3901 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3961 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4021 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4081 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4141 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4201 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4261 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4321 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4381 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4441 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4501 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa taagcggccg
     4561 cggccatgcc ggccactagt ctcgagttat tattgaagca tttatcaggg ttattgtctc
     4621 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4681 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4741 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4801 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4861 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4921 gcggcatcag agcagattgt actgagagtg caccatatga agacgtcgcc tcctcactac
     4981 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     5041 cagggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5101 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     5161 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgcccgt gtcaaggacg
     5221 gtgactgcag tgaataataa aatgtgtgtt tgtccgaaat acgcgttttg agatttctgt
     5281 cgccgactaa attcatgtcg cgcgatagtg gtgtttatcg ccgatagaga tggcgatatt
     5341 ggaaaaatcg atatttgaaa atatggcata ttgaaaatgt cgccgatgtg agtttctgtg
     5401 taactgatat cgccattttt ccaaaagtga tttttgggca tacgcgatat ctggcgatag
     5461 cgcttatatc gtttacgggg gatggcgata gacgactttg gtgacttggg cgattctgtg
     5521 tgtcgcaaat atcgcagttt cgatataggt gacagacgat atgaggctat atcgccgata
     5581 gaggcgacat caagctggca catggccaat gcatatcgat ctatacattg aatcaatatt
     5641 ggccattagc catattattc attggttata tagcataaat caatattggc tattggccat
     5701 tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac
     5761 cgccatgtt
//
