LOCUS       Exported                5673 bp ds-DNA     circular SYN 08-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5673)
  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 5673)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 8, 2024 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5673
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     regulatory      1570..1575
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1576..2142
                     /codon_start=1
                     /label=DJ-1
                     /translation="MASKRALVILAKGAEEMETVIPVDVMRRAGIKVTVAGLAGKDPVQ
                     CSRDVVICPDASLEDAKKEGPYDVVVLPGGNLGAQNLSESAAVKEILKEQENRKGLIAA
                     IAAGPTALLAHEIGFGSKVTTHPLAKDKMMNGGHYTYSENRVEKDGLILTSRGPGTSFE
                     FALAIVEALNGKEVAAQVKAPLVLKD"
     variation       1651..1653
                     /label=M26I
     variation       1765..1767
                     /label=E64D
     variation       1891..1893
                     /label=C106A
     variation       1894..1896
                     /label=A107P
     variation       2071..2073
                     /label=L166P
     polyA_signal    2261..2395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2298..2317)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2352..2371
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2418..2434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2418..2434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2431..2453)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2442..2458
                     /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(2466..2496)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2648..2665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2899..2918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3578..4438)
                     /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     4201..4220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     4648..4666
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4704..4726)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4826..4845
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4898..4909
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4913..4932)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5077..5099
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5091..5108
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5092..5108
                     /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 ccaccatggc ttccaaaaga gctctggtca tcctggctaa aggagcagag
     1621 gaaatggaga cggtcatccc tgtagatgtc atgaggcgag ctgggattaa ggtcaccgtt
     1681 gcaggcctgg ctggaaaaga cccagtacag tgtagccgtg atgtggtcat ttgtcctgat
     1741 gccagccttg aagatgcaaa aaaagaggga ccatatgatg tggtggttct accaggaggt
     1801 aatctgggcg cacagaattt atctgagtct gctgctgtga aggagatact gaaggagcag
     1861 gaaaaccgga agggcctgat agccgccatc gctgcaggtc ctactgctct gttggctcat
     1921 gaaataggtt ttggaagtaa agttacaaca caccctcttg ctaaagacaa aatgatgaat
     1981 ggaggtcatt acacctactc tgagaatcgt gtggaaaaag acggcctgat tcttacaagc
     2041 cgggggcctg ggaccagctt cgagtttgcg cttgcaattg ttgaagccct gaatggcaag
     2101 gaggtggcgg ctcaagtgaa ggctccactt gttcttaaag actaggaatt cattgatcat
     2161 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2221 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2281 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2341 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgtac
     2401 cgagctcgcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2461 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2521 taactacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2581 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2641 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3241 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3481 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3541 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3601 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3661 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3721 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3781 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3841 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3901 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3961 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4021 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4081 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4141 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4201 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4261 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4321 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4381 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4441 tcttcctttt tcaataagcg gccgcggcca tgccggccac tagtctcgag ttattattga
     4501 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4561 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4621 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     4681 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4741 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4801 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     4861 atgaagacgt cgcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     4921 tctgcataaa taaaaaaaat tagtcagggc gccattcgcc attcaggctg cgcaactgtt
     4981 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5041 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5101 cggccagtgc ccgtgtcaag gacggtgact gcagtgaata ataaaatgtg tgtttgtccg
     5161 aaatacgcgt tttgagattt ctgtcgccga ctaaattcat gtcgcgcgat agtggtgttt
     5221 atcgccgata gagatggcga tattggaaaa atcgatattt gaaaatatgg catattgaaa
     5281 atgtcgccga tgtgagtttc tgtgtaactg atatcgccat ttttccaaaa gtgatttttg
     5341 ggcatacgcg atatctggcg atagcgctta tatcgtttac gggggatggc gatagacgac
     5401 tttggtgact tgggcgattc tgtgtgtcgc aaatatcgca gtttcgatat aggtgacaga
     5461 cgatatgagg ctatatcgcc gatagaggcg acatcaagct ggcacatggc caatgcatat
     5521 cgatctatac attgaatcaa tattggccat tagccatatt attcattggt tatatagcat
     5581 aaatcaatat tggctattgg ccattgcata cgttgtatcc atatcataat atgtacattt
     5641 atattggctc atgtccaaca ttaccgccat gtt
//
